g
n
i
h
Te a c
Modern
Biology
Our systems
are used to
supplement
your
Teaching
Laboratory
MODERNBIO.COM
MODERNBIO.COM
Table of Contents
The Basic Laboratory Programs................................................................................................6
Structure, Isolation, and Function of DNA ......................................................................................6
1. Properties of DNA .............................................................................................................................................6
2. Cell Fractionation and DNA Isolation ................................................................................................................6
3. Gene Function and Cloning in Bacteria ............................................................................................................6
Animal Biology and Evolution .......................................................................................................7
1. and 2. Quantifying Amylase Activity ................................................................................................................7
3. Proteases and Factors that Influence Enzyme Activity..............................................................................................7
4. Serum Enzymes and Disease ............................................................................................................................7
5. Evolution of Serum Proteins .............................................................................................................................7
6. Identifying a Specific Enzyme in Chicken Erythrocytes by Enzyme Cytochemistry ..........................................7
Chromatographic Separation of Proteins.......................................................................................8
1. Separating Molecules by Gel Filtration.............................................................................................................8
2. The Molecular Weight of Hemoglobin..............................................................................................................8
3. Binding Specificity of Serum Albumin .............................................................................................................8
4. Properties of Amylase ......................................................................................................................................8
5. Analysis of a Protein From Egg White ...............................................................................................................8
An Introduction to Plant Cell and Molecular Biology ......................................................................9
1. Effects of Temperature on Cell Respiration .......................................................................................................9
2. Extraction and Analysis of an Enzyme from Wheat .........................................................................................9
3. Location of an Enzyme in Plant Cells and Tissues .............................................................................................9
4. Osmolarity and a Cytological Bioassay .............................................................................................................9
5. Mitotic Activity and Cell Respiration.................................................................................................................9
6. Isolating DNA from Plants ................................................................................................................................9
Introduction to Molecular Biology...............................................................................................10
101. Electrophoretic Separation of Proteins .......................................................................................................10
102. Genetics and Sickle Cell Anemia.................................................................................................................10
103. Analysis of Serum Proteins .........................................................................................................................10
104. Evolution of Serum Proteins .......................................................................................................................10
105. A Comparison of Human and Bacterial Amylase ...............................................................................................10
106. Protein Fingerprinting ....................................................................................................................... 10
Individual Experiments ............................................................................................................................. 11
IND 24. An Introduction to Electrophoresis.......................................................................................................11
IND-30 Specific Binding of Dyes to DNA .................................................................................................... 11
Tissue Specific Proteins ...............................................................................................................12
801. Serum Proteins and the Western Press-Blot...............................................................................................12
802. Identifying Ovalbumin by the Western Press-Blot ...........................................................................................12
803. Tissue-Specific Isoenzymes in the Cow ......................................................................................................12
804. Peroxidase Isoenzymes in Corn ..................................................................................................................12
Individual Experiments ............................................................................................................................. 13
IND-4. Development of the Immune System and the Western Press-Blot.........................................................13
IND-5. Specificity of Albumin Binding................................................................................................................13
Molecular Biology of Proteins .....................................................................................................14
201. Molecular Weight Determination...............................................................................................................14
202. Identifying Sex-Specific Proteins................................................................................................................14
203. Comparing Human and Bacterial Amylase ................................................................................................14
204. Peptide Mapping Analysis..........................................................................................................................14
205. Protein Evolution and the Western Blot .....................................................................................................15
206. Affinity Chromatography ...........................................................................................................................15
Individual Experiments ............................................................................................................................. 15
201P. Molecular Weight Determination ............................................................................................................16
IND-11P. Contractile Proteins from Cow Heart ..................................................................................................16
IND-25P. Isolation of Chromosomal Proteins .....................................................................................................16
Electrophoresis Package for SDS Polyacrylamide Gels .......................................................................................16
204P. Peptide Mapping Analysis ........................................................................................................................17
205P. Protein Evolution and the Western Blot ....................................................................................................17
206P. Affinity Chromatography ..........................................................................................................................17
IND-27 Composition of the Nuclesome ..............................................................................................................17
Molecular Biology of Nucleic Acids ...............................................................................................18
301. The Length of DNA Molecules ....................................................................................................................18
302. Restriction Nuclease Mapping of DNA........................................................................................................18
303. Plasmid DNA Structure ...............................................................................................................................18
304. Molecular Cloning ......................................................................................................................................18
305. Identifying Satellite Sequences ..................................................................................................................19
306. The Nucleosome Structure of Chromatin....................................................................................................19
Individual Experiments ......................................................................................................................................19
The New Genetics .......................................................................................................................20
1001. Anatomy and Evolution of the Genome ...................................................................................................20
1002. Analysis of a Genome Segment ...............................................................................................................20
1003. DNA Fingerprinting (An authentic analysis).............................................................................................20
1004. Genotype to Phenotype ...........................................................................................................................21
Individual Experiments ......................................................................................................................................21
DNA Hybridization Analysis .........................................................................................................22
401. Evolution of the Vertebrate Genome ..........................................................................................................22
402. Application of the Southern Blot Procedure...............................................................................................22
403. Detecting a Specific Sequence in the Mammalian Genome ......................................................................23
Individual Experiments ............................................................................................................................. 23
IND-16. Identifying Viral DNA by Rapid Southern Blotting ................................................................................23
Genetic Engineering ...................................................................................................................24
Tissue to Gene ............................................................................................................................26
Plant Molecular Biology..............................................................................................................27
Cloning a DNA Segment from Sheep ............................................................................................28
Mini Programs for DNA Analysis ..................................................................................................29
IND-9. Producing a Strain of E.coli that Glows in the Dark ....................................................................................29
IND-6. Analysis of a Mutant Hemoglobin Gene .................................................................................................29
IND-7. Amplification of a Hemoglobin Gene by the Polymerase Chain Reaction (PCR).....................................30
IND-10. PCR Amplification of a Gene for Ribosomal RNA from Different Organisms .........................................30
Mini Programs for DNA Analysis ..................................................................................................30
IND-21. Identifying Genomic and Plasmid DNA Sequences in E.coli by Colony PCR .........................................31
IND-12. Characterization of the Satellite DNA from the Meal Worm..................................................................31
IND-13. Transformation and Analysis of Yeast....................................................................................................32
IND-15. Bacterial Phenotype .............................................................................................................................32
IND-28. Synthetic Biology: Using Bacterial Computers to Solve the Pancake Problem......................................32
Contemporary Cell Biology .........................................................................................................33
701. Enzyme Cytochemistry ...............................................................................................................................33
702. Analysis of a Cell-Surface Receptor ............................................................................................................33
703. The Cell Nucleus..........................................................................................................................................33
Mini Programs in Cell Biology......................................................................................................34
IND-2. Tissue Printing.........................................................................................................................................34
IND-3. The ELISA Immunoassay .........................................................................................................................34
IND-22. Student Designed Research Projects: Characterization of Peroxidases in Plants ..................................35
IND-29. Electrophoretic and Chromatographic Analysis of Photosynthetic Pigments from Blue- Green Algae 35
IND-17. A Rapid Immunological Method to Study Evolution.............................................................................36
IND-14. Enzyme Kinetics ....................................................................................................................................36
Zoology Histology Developmental Anatomy ................................................................................37
IND-18. An Introduction to Zoology ...................................................................................................................37
IND-19. An Introduction to Vertebrate Histology ...............................................................................................37
IND-20. An Introduction to Developmental Anatomy........................................................................................37
IND-26. Localizing Tublin by Immunohistochemistry ........................................................................................38
The Overview Programs ..............................................................................................................39
A Molecular Approach to the An Introduction to Electrophoresis ......................................................................39
An Introduction to Molecular Genetics...............................................................................................................39
An Introduction to Electrophoresis.....................................................................................................................39
A Laboratory Course in Innovative Biology ..................................................................................40
A Laboratory Course in College-Level Biology..........................................................................42
A Laboratory Course in Cell Biology ........................................................................................43
A Laboratory Course in Molecular Biology ...............................................................................44
A Comprehensive Laboratory Course in Molecular Biology .......................................................45
A Laboratory Course in Cell and Molecular Biology ..................................................................46
A Computer Course in DNA and RNA Sequence AnalysisBiology .................................................47
Equipment ...........................................................................................................................48
Electrophoresis Equipment in the Teaching Laboratory ............................................................................. 48
Electrophoresis Equipment in the Teaching Laboratory ............................................................................. 49
Electrophoresis Equipment for the Entire Class.......................................................................................... 50
Accessories For Sample Handling .............................................................................................................. 51
Balances and Mixers.................................................................................................................................. 52
Spectrophotometer, Centrifuges and Dry Baths ........................................................................................ 53
Water Baths and Stir Plate......................................................................................................................... 54
Incubator and Shaker ................................................................................................................................ 55
Microscopes .............................................................................................................................................. 56
Thermal Cycler and Orbital Blotter ............................................................................................................ 57
Small Equipment and Labware ................................................................................................................. 58
Supplies and Expendables......................................................................................................................... 59
Individual Products and Kits for Teaching and Research ............................................................................ 60
Products for DNA Analysis ......................................................................................................................... 61
Ordering Information and Policy ............................................................................................63
Order Form ...........................................................................................................................64
On the Cover
Cover painting of DNA by Dr. Robert T. Schimke, Professor Emeritus,
Standford University, Department of Biological Sciences. Read more
about the artist on the inside back cover.
New Experiments from Modern Biology Inc.!
IND-29. Electrophoretic and Chromatographic Analysis
of Photosynthetic Pigments from Blue-Green Algae
$65.28
See Page 35 For More Information
Cyanobactera, also known as blue-green algae, obtain their
energy by photosynthesis using sunlight as their energy source.
These organisms have been considered to be the oldest and
the most important bacteria on the earth. It is believed that
they were responsible for the initial oxygenation of the earth's
atmosphere through photosynthesis and it is also felt they were
the precursors to the chloroplasts that are found in true algae and
plants. In part A of this exercise, students prepare a water-soluble
extract from blue green algae and show that it contains the
single major protein Phycocyanin by electrophoresis as shown
in the gel below. They also determine the charge
of this protein by comparing its electrophoretic IND-30. Specific Binding of Dyes to DNA
mobility to the mobilities of dyes with known
Proteins that bind to DNA control the
charges. In part B, they prepare an alcohol extract
processes of gene regulation and DNA
and analyze the smaller alcohol soluble pigments by
replication. The electrophoretic mobility
thin layer chromatography in order to identify the
band shift assay is a common technique used
chlorophylls and major carotenoid pigments. The
to study these proteins and to study specific
results of this two-part study give students practical
protein-DNA interactions. In this exercise,
hands-on experience with isolation of components
students use this assay to identify dye
from cells as well as electrophoresis and thin layer
molecules that bind to DNA and attempt to
chromatography and introduces them to one of the
determine the mechanism by which these
most important organisms on the earth.
drugs interact with the DNA molecule.
$19.99
Equipment Packages!
PIPETTOR PACKAGE
$378.00
!
w
o
N
r
Orde
For the Value of $512.00
1 Pipettor Stand & 4 Pipettors of your choice
Mechanical Adjustable Volume Pipettor
Our mechanical adjustable pipettors offer accurate and precise sampling and dispensing of liquid volumes. The
pipettors come in a variety of ranges to meet your needs. All pipettors have been quality tested according to ISO
8655/DIN 12650 and come with a one year warranty.
Features:
t Easy to calibrate and maintain
t Ejector collar and tip cone can be removed for easy cleaning and maintenance
t Digital display clearly reads volume setting
t Supplied with full documentation, warranties and certificates
CAT NO.
720080
720020
720070
720060
CAPACITY
2-20ul
5-50µl
20-200µl
100 - 1000µl
PRICE
$118
$118
$118
$118
Micropette™
Pipettor Stand
Linear Pipettor Stand, holds up to 6 pipettors.
Item No. 71000085
Reg. Price $40.00
See Page 51 For More Information
s&!8s7EBSITEWWWMODERNBIOCOMs
1
4
Suitability for your Teaching Laboratory
Be
gin
In ning
ter
m
A d ed i
va
a
nc te
ed
Level
1. Individual Experiments
Recommended
Ce
ll
B
Mo iolo
lec gy
Ge ular
ne
Bi
t
olo
Mi ics
gy
cr
ob
An iol
im og
y
a
P l l St
an
ud
t
ie
Ev Stud s
olu ie
s
Hu tion
ma / I
m
Stu n Di mu
de sea no
log
nt
s
D e es
y
sig
ne
d
Highly Recommended
Topic
Page
No Electrophoresis
Properties of DNA
Cell Fractionation and DNA Isolation
Gene Function and Cloning in Bacteria
Evolution of Serum Proteins
Effects of Temperature on Cell Respiration
Analysis of an Enzyme from Wheat
Producing E. coli that Glows in the Dark
Transformation and Analysis of Yeast
Bacterial Phenotypes
Digestive Enzymes
Analysis of a Cell-Surface Receptor
The Cell Nucleus
Tissue Printing
The ELISA Immunoassay
Immunological Methods and Evolution
Enzyme Kinetics
Zoology / Histology
Developmental Anatomy
Enzyme Cytochemistry
Analysis of a Cell-Surface Receptor
DNA and RNA Sequence Analysis
Evolution of the Vertebrate Genome
Localizing Tubulin by Immunohistochemistry
Non-Denaturing
Protein
Electrophoresis
Non-Denaturing Protein
Electrophoresis
(Electrophoresis
Package
- Page
(Electrophoresis Package
1/81/8
- Page
11) )
Electrophoretic Separation of Proteins
Genetics and Sickle Cell Anemia
Analysis of Serum Proteins
Binding of Dyes to DNA
Human and Bacterial Amylase
Protein Fingerprinting
Pigments in Blue Green Algae
Protein Evolution and the Western Blot
Tissue-Specific Isoenzymes in the Cow
Peroxidase Isoenzymes in Corn
Immune System and the Western Blot
Specificity of Albumin Binding
SDS-Denaturing
Protein
Electrophoresis
SDS-Denaturing
Protein
Electrophoresis
(Electrophoresis
Package
(Electrophoresis Package
2 & 22M&-2M
Page- Page
15) )
Molecular Weight Determination
Human and Bacterial Amylase
Peptide Mapping Analysis
Protein Evolution and the Western Blot
Affinity Chromatography
Contractile Proteins from Cow Heart
Isolation of Chromosomal Proteins
s-ODERN"IOLOGY)NCss
6
6
6
7
9
9
29
32
32
7
33
33
34
34
36
36
37
37
33
33
47
22
38
10
10
10
11
10
10
35
12
12
12
13
13
14/16
14
14/17
14/17
15/17
16
16/17
5
Suitability for your Teaching Laboratory
DNAElectrophoresis
Electrophoresis
DNA
(Electrophoresis
Package
- Page
(Electrophoresis Package
3/43/4
- Page
19) )
The Length of DNA Molecules
Restriction Nuclease Mapping of DNA
Plasmid DNA Structure
Molecular Cloning
Identifying Satellite Sequences
Nucleosome Structure of Chromatin
Anatomy and Evolution of the Genome
Analysis of a Genome Segment
DNA Fingerprinting
Genotype to Phenotype
Analysis of a Mutant Hemoglobin Gene
Amplification of a Hemoglobin Gene by PCR
PCR Amplification of Ribosomal RNA Gene
Identifying Sequences in E.coli by Colony PCR
Satellite DNA from the Meal Worm
Application of the Southern Blot Procedure
A DNA Sequence in the Mammalian Genome
Identifying Viral DNA by Rapid Southern Blotting
Be
gin
In ning
ter
m
A d ed i
va
a
nc te
ed
Level
Recommended
Ce
ll
B
Mo iolo
lec gy
Ge ular
ne
Bi
t
olo
Mi ics
gy
cr
ob
An iol
im og
y
a
P l l St
an
ud
t
ie
Ev Stud s
olu ie
s
Hu tion
ma / I
m
Stu n Di mu
n
s
de
nt ease olog
De s
y
sig
ne
d
Highly Recommended
Topic
Page
18
18
18
18
19
17/19
20
20
20
21
29
30
30
31
31
22
23
23
2. Experiments with a Related Theme
Structure, Isolation, and Function of DNA
Animal Biology and Evolution
Chromatographic Separation of Proteins
Plant Cell and Molecular Biology
Introduction to Molecular Biology
Tissue-Specific Proteins
Genetic Engineering
Tissue to Gene
Molecular Biology of Proteins
Molecular Biology of Nucleic Acids
Plant Molecular Biology
Cloning a DNA Segment
Peroxidases in Plants
DNA Hybridization Analysis
The New Genetics
Contemporary Cell Biology
Human Genetics and Disease
Introduction to Molecular Genetics
Introduction to Electrophoresis
Synthetic Biology
6
7
8
9
10
12
24
26
14
18
27
28
35
22
20
33
39
39
39
32
3. College-Level Laboratory Courses
General Biology
An Introduction to Cell and Molecular Biology
Cell Biology
Molecular Biology
Comprehensive Molecular Biology
A Laboratory Course in Investigative Biology
s&!8s7EBSITEWWWMODERNBIOCOMs
42
46
43
44
45
40
BASIC PROGRAMS
6
The Basic Laboratory Programs
GENERAL FEATURES
The four basic programs are supplied complete with all of
the laboratory manuals, chemicals, and accessories needed
for beginning students to perform exciting experiments in
modern biology. Electrophoresis equipment is not required
in all four of the basic programs.
All programs are designed to be used by 16 students working
in pairs. In most cases, each experiment can be completed
within a 90 minute laboratory session.
The programs are intended to introduce advanced secondary
and beginning college students to contemporary topics
including enzyme action, DNA structure and function,
molecular evolution, and DNA cloning.
A full-semester course in college-level biology was created
from experiments in the programs and is described on page
43 of this catalog.
Structure, Isolation, and Function of DNA
Basic Laboratory Program 1
This program is used to explore some of the major concepts and techniques that have revolutionized the biological sciences
during the past two decades. Without the use of hazardous chemicals, students obtain an introduction and practical experience
with the structure and isolation of DNA, cell fractionation, and gene cloning procedures.
EXPERIMENTS
1. Properties of DNA
The DNA molecule from a single
human chromosome is about 4 cm
long and the length of DNA in an
individual is about 200 times the
distance from the earth to the sun.
Isolated DNA in a test tube is also a
long, stiff molecule. When alcohol
is added to a DNA solution, the DNA
fibers precipitate and can be spooled
onto a glass rod. This feature of DNA is illustrated in the exercise, which
provides enough purified DNA for 16 students working in pairs to
perform the experiment. The spooling phenomenon is also used by the
student to study the double-stranded nature of DNA and to investigate
the effect of breaking DNA into small pieces with the enzyme DNAse.
Ethyl alcohol (90-100%) is needed but not provided.
2. Cell Fractionation and DNA Isolation
Students isolate nuclei from calf thymus tissue and examine them
microscopically. The DNA is then extracted from the nuclei by a simple
procedure that uses a detergent and alcohol. Microscopes and a small
centrifuge are desirable but not absolutely necessary for the exercise.
3. Gene Function and Cloning in Bacteria
Occasionally, E.coli cells are found in nature that are resistant to the
toxic effects of the antibiotic ampicillin. In this exercise, students
create such an ampicillin-resistant population of E.coli by introducing
into bacterial cells a plasmid that contains an ampicillin-resistance
gene. Sufficient sterile materials are supplied for sixteen platings. The
introduction of plasmid DNA into bacterial cells is called transformation
and has enabled scientists to obtain large quantities of more than
1000 genes including those for human interferon, insulin, and growth
hormone. This exercise requires minimal teacher preparation and
provides a laboratory experience with
A waterbath is optional.
one of the most important techniques
See page 54.
used in gene-cloning studies.
Materials Provided
*19
*20
*24
2
*30
10
Sterile Tubes
Petri Dishes
Inoculating Loops
Macropipetors
Sterile Transfer Pipets
Small Transfer Pipets
8 Tube Racks
8 Glass Vials
8 Glass Rods
Nuclear Buffer
*Plasmid DNA
Cheese Cloth
Calf Thymus DNA
DNAse I
Calf Thymus Tissue
*Provided sterile and ready to use
CAT. NO.
Nuclear Stain
Sodium Dodecyl
Sulfate
*Nutrient AgarAmpicillin
*Nutrient Broth
*CaCl2
E.coli
Ampicillin
DESCRIPTION
Price List - Basic Laboratory Program 1
B1
B1-BK
B1-SM
The Chemical Package for 16 students
working in pairs plus one student and
one instructor manual (The student manual
may be reproduced for educational purposes).
8 Student Manuals
1 Student Manual and 1 Instructor Manual
PRICE
$149.99
$48.12
$6.87
Single Experiments Offered From Basic Program 1
B1-1
B1-1F
B1-2
B1-3
Properties of DNA
Chemical refill for (B1-1) Properties of DNA
Contains DNA solution and DNAse
Cell Fractionation and DNA Isolation
Gene Function and Cloning in Bacteria
$54.48
$35.55
$54.48
$73.34
Multi Section Lab Experiments offered from Basic Program 1
Each MSL (Multi Section Lab) experiment provides one set of reusable items and four sets
of chemicals which can be used to teach 4 Sections of 16 students working in pairs.
B1-2 MSL Cell Fractionation and DNA Isolation
s-ODERN"IOLOGY)NCss
$107.00
7
Basic Laboratory Program 2
This collection of experiments was prepared for the novice to help bridge the gap between traditional animal physiology and the
biochemistry of cell processes in animals. Following the step-by-step procedures in their laboratory manuals, students will study
a variety of topics including the action, properties, and cell location of enzymes, and the evolution of mammalian proteins.
A centrifuge is helpful
but not necessary
EXPERIMENTS
1. and 2. Quantifying Amylase Activity
This exercise begins with a discussion of
the digestive enzymes produced by the
salivary glands, stomach, intestine and
pancreas. The digestive system of a rodent
is provided to familiarize students with
anatomical features of the digestive organs
of a mammal. Students are then given the
opportunity to prepare enzyme extracts from pancreatic tissue
and to determine the amount of amylase in the extracts and in
their own saliva. This exercise offers practical experience with
enzyme extraction procedures and introduces the concept of the
calibration curve for determining the levels of a specific enzyme
in a complex mixture.
In the figure above, starch was incorporated into an agar gel, and
samples containing amylase were placed in sample wells made in
the agar. After one day, the starch was stained with iodine. Areas
where the starch was degraded by amylase are seen as clear rings
around the wells and the diameters of the rings are proportional
to the concentration of amylase.
3. Proteases and Factors that Influence Enzyme Activity
Students contrast the effects of pH on the activities of gastric pepsin
and pancreatic chymotrypsin and relate their results to the acidic
pH of the stomach and neutral pH of the small intestine. The effects
of temperature and thermal denaturation on enzyme activity are
also investigated.
4. Serum Enzymes and Disease
Tissue injury often results in the leakage of large amounts of cellular
enzymes into the circulatory system and measurements of the
amounts and types of enzymes in serum are frequently made for
the diagnosis of human disease. For example, inflammation of the
pancreas is accompanied by the release of pancreatic amylase and
proteases into the blood stream. In this exercise, students determine
the levels of amylase and proteases in serum and compare these
levels to those found in saliva and pancreatic extracts.
This concept is illustrated in this exercise where students examine
the ability of antibodies against cow serum proteins to react
with serum proteins from cow, sheep, goat, horse, donkey, and
chicken. The results of this study show that sheep and goat are
most closely related to cow which is in agreement with classical
taxonomy.
6. Identifying a Specific Enzyme in Chicken Erythrocytes by
Enzyme Cytochemistry
The concept that different enzymes are found in different regions
of a cell is illustrated in this exercise. Here, students use the
technique of enzyme cytochemistry to show that the enzyme lactate
dehydrogenase is located in the cytoplasm of chicken erythrocytes.
The exercise introduces students to the basic structure of animal
cells and provides an illustration of an important technique used in
modern cell biology. Eight chicken blood smears are provided for
the experiment and microscopes are required.
Chemical Package Contents
8
50
60
20
2
16
20
8
Tube Racks
Small Test Tubes
Transfer Pipets
Petri Dishes
Macropipetors
Large Test Tubes
Pasteur Pipets
Chicken Blood
Smears
3 Plastic trays/lids
Methylene Blue
Eosin
CAT. NO.
B2
Digestive System
(from rat or mouse)
Pepsin
Extraction Buffer
Pancreatic Powder
Amylase
Pancreatic Extract
(4 tubes)
Starch Solution
Agar
Gel Buffer
LDH Substrate
Potassium Iodine
Casein Solution
Chymotrypsin
Chicken Serum
Horse Serum (2 tubes)
Cow Serum Antibodies
Cow Serum
Sheep Serum
Goat Serum
Donkey Serum
NP-40
DESCRIPTION
Price List - Basic Laboratory Program 2
PRICE
$199.80
B2-BK
The Chemical Package for 16 students
working in pairs plus one student and
one instructor manual (The student manual
may be reproduced for educational purposes).
8 Student Manuals
B2-SM
B2-CR
1 Student Manual and 1 Instructor Manual
Chemical Refill Package for Basic Program 2
$6.87
$149.77
$48.12
5. Evolution of Serum Proteins
Each protein carries in its amino acid sequence information
Single Experiments Offered From Basic Program 2
B2-5
Evolution of Serum Proteins
$56.79
pertaining to its evolutionary history and origin, and provides
(Previous
catalog
number
was
B2-4)
clues to the evolutionary history of the organism in which it is
found. Indeed, proteins existing today are in effect living fossils.
s&!8s7EBSITEWWWMODERNBIOCOMs
BASIC PROGRAMS
Animal Biology and Evolution
BASIC PROGRAMS
8
Chromatographic Separation of Proteins
Basic Laboratory Program 3
Gel filtration is a chromatographic method used to
separate molecules of different sizes. This technique
is an important tool for isolating and characterizing
proteins. In this series of experiments, students use this
method along with basic enzyme and antibody assays to
study the structure and function of proteins.
5. Analysis of a Protein From Egg White
Students first isolate ovalbumin, a major protein found in egg
white. They then use gel filtration and a simple immunological
procedure to determine the size of this protein and the
amount of it in the egg.
Materials Provided
EXPERIMENTS
1. Separating Molecules by Gel Filtration
Students fill their chromatographic columns with packing
material and separate three colored molecules of known
molecular weights as shown
in the photograph. The
exercise illustrates how
components of a complex
mixture can be separated
by chromatography and
introduces the concept
of the standard curve
for determining the size
of an unknown protein.
Watch a video of this experiment in
action at MODERNBIO.COM
2. The Molecular Weight of Hemoglobin
Students determine the size of hemoglobin and estimate the
number of amino acid residues in this protein.
3. Binding Specificity of Serum Albumin
The binding of an enzyme to its substrate is only one example
of the many specific molecular interactions that occur in
biological systems. An analogous binding process occurs
with serum albumin which binds certain small molecular
weight molecules in blood. In this exercise, students use a
gel filtration assay to examine the binding of various dyes to
albumin and determine if the native shape of the protein is
required for its binding specificity.
4. Properties of Amylase
Gel filtration chromatography has the advantage of being able
to yield the size of different proteins in a complex mixture.
This feature is illustrated in this experiment where students
compare the molecular weight of amylase from pancreas and
saliva.
Chemical Package Contents
8 Collection Plates
70 Small Test Tubes
60 Transfer Pipets
20 Petri Dishesand
2 Macropipetors
16 Large Test Tubes
20 Pasteur Pipets
8 Tube racks
Cheese cloth
SDS
CAT. NO.
8 Chromatographic
Columns (column tubes,
reservoirs, lower fittings,
bed supports caps)
Column Packing Material
Column Buffer
Phenol Red
Myoglobin-Blue Dextran
Cytochrome
C-Hemoglobin
Lysis Buffer
Rabbit Blood
Bromophenol Blue
Cow Serum Albumin(2)
Agar
Starch Solution
Potassium Iodine
Amylase
Pancreatic Extract
Ovalbumin
Ovalbumin Antibodies
DESCRIPTION
Price List - Basic Laboratory Program 3
B3
B3-BK
B3-SM
B3-CR
The Chemical Package for 16 students
working in pairs plus one student manual and
one instructor manual (The student manual may
be reproduced for educational purposes.)
8 Student Manuals
Sample Student Manual (43 pages) plus one
instructor manual.
Chemical Refill Package for Basic Program 3
PRICE
$216.31
$48.12
$6.87
$152.73
Single Experiments Offered From Basic Program 4
B3,1-3
Mini Program in Chromatography
$127.90
Includes chemicals and instructions for experiments 1-3.
This series can be completed in two
2-3 hour laboratory sessions.
s-ODERN"IOLOGY)NCss
9
Basic Laboratory Program 4
Recent discoveries in the plant sciences have illustrated the potential of modern techniques for the production of genetically
engineered plants that are resistant to pathogens, herbicides, and adverse environmental conditions. A solid foundation in basic
plant cell and molecular biology is required in order for students to participate in this green revolution. The novel experiments in
this program are intended to provide this background. In the program, students extract and characterize enzymes and DNA from
plant tissues, localize enzymes in plants at the tissue and cell levels, and analyze fundamental process common to all cells including
mitosis, osmosis, and cell respiration. The program is ideal for introducing the beginning student to basic biological principles
and plant biology and can serve as a supplement to elementary laboratory courses in botany, plant physiology, and agriculture.
Microscopes, slides, cover slips, a tabletop centrifuge, ethyl alcohol, and hydrochloric acid are needed, but not provided.
1. Effects of Temperature on Cell Respiration
Cell respiration can be viewed as a series of enzyme catalyzed
reactions in which carbohydrates, proteins, and fats are broken down
to carbon dioxide and water with the release of energy. During the
process, hydrogen is removed from the fuel molecules and oxygen is
consumed. With this background information, students measure oxygen
consumption and hydrogen liberation in germinating barley at different
temperatures. The program provides eight calibrated respirometers for
measurement of oxygen consumption and the chemicals required to
perform a graphic dye reduction assay. The exercise introduces students
to a fundamental biological process and provides insight into seed
structure and germination.
2. Extraction and Analysis of an Enzyme from Wheat
Acid phosphatase is present in many plant tissues where it catalyzes
the removal of phosphate groups from macromolecules at low pH.
In this exercise, students prepare a cell-free extract from wheat
germ and determine the amount of the enzyme present in the extract.
The experiment offers practical experience with enzyme extraction
procedures and is an excellent introduction to the analysis of enzyme
activity and basic enzyme kinetics. A colorimeter is desirable but not
absolutely necessary for this exercise.
3. Location of an Enzyme in Plant Cells and Tissues
Flowering plants are multicellular organisms and different enzymes are
frequently produced in different tissues and in different cell types. This
concept is illustrated in the first part of the exercise where students use an
enzyme histochemical procedure to show that peroxidase is produced by
cells that comprise a layer in the corn seed called the aleurone as shown
in the figure below. In the second part of the exercise, a similar procedure
is used by the student to show that peroxidase is found in the cell wall of
onion epithelial cells.
The picture shows typical results of the first
portion of Experiment 3. Germinating corn seeds
were incubated in water, iodine, tetrazolium, or
in a peroxidase substrate. Note that the triangular
embryo exhibits a high rate of respiration, the
aleurone layer contains peroxidase and the starchy
endosperm is stained with iodine. Students also
locatlities the site of cellular respiration and starch
storage as shown.
4. Osmolarity and a Cytological Bioassay
In this exercise, osmosis is examined in living onion epithelial cells
which are used in a cytological assay to determine the osmolarity of an
unknown sucrose solution provided with the program.
5. Mitotic Activity and Cell Respiration
The onion root has served as a model system for the study of mitosis,
cell elongation, and cell differentiation in plants. In this experiment,
students examine various segments of onion roots for cytological
differences and then correlate their cytological findings with different
respiration rates in the root segments. The exercise provides practical
experience with tissue fixing, staining and slide making procedures, and
introduces students to the concepts of cell division, the cell cycle, and
differentiation in plant systems.
6. Isolating DNA from Plants
A first step in plant recombinant DNA research frequently involves the
isolation of DNA from plant tissue. Students perform this procedure
in the exercise where they prepare DNA from wheat germ by a simple
procedure that uses a detergent, salt, and alcohol.
Chemical Package Contents
Calibrated Respirometers
Collection Plates
Glass Rods
Macropipets
Small Transfer Pipets
Large Transfer Pipets
Tetrazolium
Barley Seeds
Potassium Hydroxide
Corn Seeds
Onion Bulbs
CAT. NO.
B4-BK
B4-SM
B4-CR
B4-2
DESCRIPTION
PRICE
Price List - Basic Laboratory Program 4
B4
B4-1
Hydrogen Peroxide (2 tubes) Methylene Blue
Potassium Iodine
Eosin
EDTA
Sodium Dodecyl
Sulfate
Tris Buffer (2 tubes)
Ammonium Acetate
Chloronapthol
Acid Phosphatase
Wheat Germ
Substrate
Acid Phosphatase
Nitrophenol Standard
Extraction Buffer
Large Test Tubes
Sucrose Solution
Cheese Cloth
The Chemical Package for 16 students working in $193.12
pairs plus one student and one instructor manual.
8 Student Manuals
$48.12
Sample Student Manual plus one instructor manual. $6.87
Chemical Refill Package for Basic Program 4
$152.73
Single Experiments Offered From Basic Program 4
Effects of Temperature on Cell Respiration
Extraction and Analysis of an Enzyme from Wheat
$64.79
$63.73
Multi Section Lab Experiments offered from Basic Program 1
Each MSL (Multi Section Lab) experiment provides one set of reusable items and four sets
of chemicals which can be used to teach 4 Sections of 16 students working in pairs.
B4-1 MSL Effects of Temperature on Cell Respiration
B4-2 MSL Extraction and Analysis of an Enzyme from Wheat
s&!8s7EBSITEWWWMODERNBIOCOMs
$120.06
$117.47
BASIC PROGRAMS
An Introduction to Plant Cell and Molecular Biology
10
Introduction to Molecular Biology
Standard Laboratory Program 1
PRO
ELECTR TEIN
OPHO
-NATIV RESIS
E-
This program stresses the relevance of molecular biology to the study of physiology and human disease. Students first learn how
proteins are separated on the basis of charge by electrophoresis. With this information, they perform a series of experiments
aimed at illustrating the importance of the technique in biology and medicine. Students examine proteins in biological fluids
and relate their findings to physiological and pathological states. They also study specific antigens and enzymes in serum
and the abnormal hemoglobin in patients suffering from sickle cell anemia. This fine collection of experiments provides an
introduction to electrophoresis and to protein composition, structure, and function in health and disease.
PROTIENS
101. Electrophoretic Separation of Proteins
EXPERIMENTS
Students are introduced to the theory of separating proteins according
to charge differences using electrophoresis. They then study four
proteins and relate differences in their charges to their migration rates
in an electric field. Each protein is a different color so that its progress
Staining is not necessary.
during the separation can easily be followed.
102. Genetics and Sickle Cell
Anemia
Watch this experiment in
action: MODERNBIO.COM
Normal and Sickle Hemoglobin
EXP 102
Many changes in the structure of
1 2 3 4 5 6 7 8
hemoglobin have arisen by mutations.
About one person in 100 carries a mutant
hemoglobin gene, and these individuals
have abnormal hemoglobin molecules
in their blood. One of the most common
abnormal hemoglobins is hemoglobin S,
which causes sickle cell anemia. When
the gene for hemoglobin S is inherited
from both parents, all of the hemoglobin
in the circulation is hemoglobin S
and the individual suffers from severe Inthisanalysis,hemoglobin
from a normal individual
anemia. When the gene for hemoglobin (gel lanes 1, 4, 5, 8) is
S is inherited from only one parent, the compared to hemoglobin
individual is heterozygous for the condition S (lanes 3 and 7) and
and has sickle cell trait. Although these hemoglobin from a person
individuals rarely have severe anemia, who has sickle cell trait
(lanes 2, and 6). Note the
half of their hemoglobin is hemoglobin two forms of hemoglobin
S and half is normal hemoglobin. In in the individual with sickle
this exercise, students compare the cell trait.
electrophoretic patterns of hemoglobin
from a normal individual to hemoglobin S and hemoglobin from a person
with sickle cell trait. Students are also given the opportunity to study their
own hemoglobin in order to test for hemoglobin variants. Typical results
of this exercise are shown above.
103. Analysis of Serum Proteins
Serum contains a variety of small molecules, as well as hundreds of
different serum proteins. Serum proteins are frequently separated
and characterized by electrophoresis in the clinical laboratory to
determine the concentration of various proteins and to detect abnormal
protein species. In this exercise, students use electrophoresis to study
cow serum and to identify major serum proteins including albumin,
transferrin and antibodies. This exercise serves as a good introduction
to the function and complexity of proteins in blood.
104. Evolution of Serum Proteins
Antibodies can be used to study evolution at the molecular level. In
this exercise, immuno-electrophoresis and double diffusion are used
to illustrate this important concept. The analysis shows that cow serum
proteins are similar to those from sheep and goat but are distinct from
serum proteins from horse, donkey and chicken. This exercise acquaints
students with basic immunological techniques and serves as an excellent
illustration of the utility of these techniques for the study of protein
evolution. This exercise requires two 2-3 hour laboratory periods.
105. A Comparison of Human and Bacterial Amylase
A powerful technique used in the modern laboratory involves the
detection of specific enzymes in agarose gels following electrophoresis of
proteins from serum, tissue extracts and other biological preparations.
This procedure makes it possible to study certain properties of
individual enzymes in complex protein mixtures. In this exercise,
students are introduced to this procedure by comparing the amylase
found in bacteria with that in their own saliva.
Extra LDH can be purchased
to expand this Experiment.
See page 60 CAT. 1-10.
106. Protein Fingerprinting
A comparison of specific proteins from
different species provides a powerful
approach for establishing evolutionary
relationships and for identifying organisms.
A common approach used for this purpose is
protein fingerprinting where electrophoretic
properties of specific proteins are analyzed
in different species. In this exercise, students
use the approach to compare the forms of
the enzyme lactate dehydrogenase that are
found in serum of different mammals. The
photo shows typical results of this graphic
experiment. The results illustrate that each
species has a characteristic pattern of bands
and that the pattern in sheep and goat is
similar, while the patterns from cow and
especially horse are distinct.
Forms of lactate dehydrogenase
in serum - EXP 106
1
2 3 4 5 6 7 8
Serum proteins from cow
(lanes 1, 5), sheep (lanes
2, 6), goat (lanes 3, 7), and
horse (lanes 4, 8) were
separated by electrophoresis.
The different forms of lactate
dehydrogenase were then
detected by activity staining.
CAT. NO.
DESCRIPTION
PRICE
S1
The Chemical Package for 16 students
$202.17
working in pairs plus 1 student and 1 instructor manual.
S1-BK 8 Student Manuals
$63.14
S1-SM Sample Student Manual plus one instructor manual. $9.61
s-ODERN"IOLOGY)NCss
11
INDIVIDUAL EXPERIMENTS
Select the individual experiments in this series that best suit
your needs. Each experiment is supplied with the chemicals
needed for 16 students working in pairs. Each one also comes
with an instructor guide and a single student guide which you
may reproduce for educational purposes. Student guides give
background information and detailed instructions that are
necessary for successful performance and comprehension of
the experiments. The instructor guide illustrates the results of
the exercise and gives answers to the study questions posed in
the student guide. If you chose one or more of the experiments
Electrophoresis Package 1/8
(For up to 6 individual experiments)
$62.42
Single Experiments from Standard Program 1
The following experiments include Chemicals and Instructions.
EXP-101 Electrophoretic Separation of Proteins
$42.44
EXP-102 Genetics and Sickle Cell Anemia
$46.54
EXP-103 Analysis of Serum Proteins
$48.86
EXP-104 Evolution of Serum Proteins
$71.77
EXP-105 A Comparison of Human and Bacterial Amylase
$47.92
EXP-106 Protein Fingerprinting
$59.89
IND-24 and Experiment-101 are each in
overview program 3 see page 39.
Save over $42 by purchasing 03.
IND 24. AN INTRODUCTION TO ELECTROPHORESIS
Part A. Identifying Unknowns by electrophoresis
Part B. Identifying Dyes that Bind to DNA
This economical 2-part exercise provides an introduction to
agarose gel electrophoresis. In
the first part of the exercise,
students identify unknown dye
molecules by comparing their
electrophoretic migration with
the migration of known dyes. In
the second part, students identify
dye molecules that bind to DNA
and determine the mechanism.
Chemical Package Contents
EP-1/8
Agarose
Gel Stain (Coomassie blue)
Electrophoresis Buffer
Hemoglobin Elect. Buffer
EXP-101
Cytochrome C
Hemoglobin
Serum Albumin
Myoglobin
EXP-102
Normal Human Hemoglobin
Sickle Hemoglobin
Sickle Trait Hemoglobin
Erythrocyte Lysis Buffer (2)
EXP-103
Cow Serum
Cow Gamma Globulins
Cow Transferrin
Cow Serum Albumin
EXP-104
Donkey Serum
Goat Serum
Cow Serum
Horse Serum
Sheep Serum
Chicken Serum
Filter Paper
Pasteur Pipets
Cow Serum Antibodies
Petri Dishes
Tris Buffer Saline
EXP-105
Starch Solution
Potassium Iodine Solution
Sample Buffer
Alpha-amylase
PROTIENS
EP-l/8
below, you should also order Electrophoresis Package 1/8. This
Electrophoresis Package provides sufficient agarose, gel stain and
electrophoresis buffers for up to 6 of the individual experiments
in this series.
EXP-106
LDH-Substrate
Cow Serum
Sheep Serum
Goat Serum
Horse Serum
Electrophoresis Sample Buffer
The exercise was designed for 8 groups of students and includes
four colorful dye mixtures and DNA. Electrophoresis Package 1/8
is needed but not provided.
CAT. NO.
DESCRIPTION
IND-24 An Introduction to Electrophoresis
PRICE
$19.99
IND30 SPECIFIC BINDING OF DYES TO DNA
Proteins that bind to DNA control the processes of gene regulation
and DNA replication. The electrophoretic mobility band shift
assay is a common technique used to study these proteins and
to study specific protein-DNA interactions. In this exercise,
students use this assay to identify dye molecules that bind to DNA
and attempt to determine the mechanism by which these drugs
interact with the DNA molecule.
CAT. NO.
DESCRIPTION
IND-30 Specific Binding of Dyes to DNA
s&!8s7EBSITEWWWMODERNBIOCOMs
PRICE
$19.99
12
PRO
ELECTR TEIN
OPHO
-NATIV RESIS
E-
Tissue Specific Proteins
Standard Laboratory Program 8
Electrophoresis is a powerful tool for separating proteins in complex mixtures. However, it is often necessary to identify specific
enzymes and structural proteins after electrophoresis so they can be studied in further detail. In this series of experiments,
students use electrophoresis in conjunction with novel immunological and enzymatic procedures to study tissue-specific proteins
in animals and plants. As in all of our programs, enough materials for 8 groups of students are included. Sufficient materials are
provided so that experiments 803 and 804 can be preformed twice: once with the tissues provided with the program and once
with tissues of your choice. The PROCELL Electrophoresis Unit, a tabletop centrifuge, and a 37°C waterbath are required.
801. Serum Proteins and the Western Press-Blot
Western blotting is one of the most powerful methods in molecular
biology for identifying and characterizing specific proteins in
complex protein mixtures. We have now streamlined westernblotting procedures so that the entire analysis can be performed
during a single 3-hour, or two 2-hour laboratory sessions. In
exercise 801, students use the procedure outlined below to identify
albumin, transferrin and gamma globulins in serum and then to
study the evolutionary relationships of albumin in vertebrates.
802. Identifying Ovalbumin by the Western Press-Blot
Chicken egg white contains a variety of proteins including
ovalbumin, conalbumin, ovomucoid and lysozyme. In this exercise,
students use the western press-blot procedure to characterize
chicken ovalbumin and then to determine if the protein is found
in egg whites from turkey and quail.
803. Tissue-Specific Isoenzymes in the Cow
Isoenzymes are different molecular forms of the same enzyme and
five major lactate dehydrogenase (LDH) isoenzymes are found in
vertebrate tissues. The amounts of the isoenzymes vary in a tissue
specific manner and these differences can be readily detected by
localizing LDH activity in an agarose gel after electrophoresis of
tissue extracts. In this exercise, students prepare a tissue extract
from calf thymus and then compare the LDH isoenzyme profile to
those from calf serum, heart and muscle.
804. Peroxidase Isoenzymes in Corn
Peroxidase is associated with the plant cell wall and different
forms of peroxidase are found in different plant tissues. These
tissue-specific differences are revealed by this exercise where
students examine the electrophoretic patterns of peroxidase in
corn root and shoot extracts.
Peroxidase Isoenzymes EXP-804
Negative Electrode
<
Corn extracts and peroxidase
standards were separated
by electrophoresis and the
different peroxidase isoenzymes were then detected
by activity staining. A simple
procedure is described for the
exercise which enables the
student to position the sample
wells in the middle of the gel.
This operation permits the
detection of both acidic and
basic forms of peroxidase.
<
PROTIENS
EXPERIMENTS
Positive Electrode
s-ODERN"IOLOGY)NCss
13
Chemical Package Contents
Price List - Standard Program 8
CAT. NO.
DESCRIPTION
PRICE
S8
The Chemical Package for 16 students working
$235.14
in pairs plus student manual and one instructor manual
(The student manual may be reproduced for educational purposes.)
S8-BK
S8-SM
8 Student Manuals
$63.14
Sample Student Manual plus one instructor manual. $9.61
INDIVIDUAL EXPERIMENTS
EP-1/8
EXP-801
EXP-802
EXP-803
EXP-804
Electrophoresis Package 1/8
Serum Proteins and the Western Press-Blot
Identifying Ovalbumin by the Western Press-Blot
Tissue-Specific Isoenzymes in the Cow
Peroxidase Isoenzymes in Corn
$ 62.42
$ 96.41
$ 96.41
$ 74.87
$ 68.09
IND4. DEVELOPMENT OF THE IMMUNE SYSTEM
AND THE WESTERN PRESSBLOT.
In most mammals, antibody production does not occur until after
birth. The newborn calf receives antibodies in milk and these
antibodies are responsible for passive immunity during early
postnatal life. Thus, fetal calf serum is devoid of antibodies, neonatal
calf serum contains antibodies of maternal origin while serum from
the adult contains large amounts of antibodies made by the mature
immune system. This fascinating developmental scheme is illustrated
in this exercise. First, students electrophorese serum proteins
from fetal, newborn, and adult animals on an agarose gel and the
separated proteins are transferred onto a nitrocellulose membrane
by the Western Press-Blot procedure shown on the facing page. All
proteins on the blot are stained with protein blot stain and then
the major class of circulating antibodies (IgG) is detected by using
an immunological procedure. The results show that albumin and
transferrin are present in sera from all stages while the antibodies
are absent in fetal serum but present at high levels in serum from the
adult. This analysis can be performed during a single 3-hour, or two
2-hour laboratory sessions. The exercise was designed for 8 groups
of students and includes: fetal calf serum, newborn calf serum,
adult cow serum, anti-Cow IgG (peroxidase linked), nitrocellulose,
blotting paper, protein blot stain, Tris buffer saline (TBS), TBS +
NP40, color development buffer, chloronapthol, cow transferrin,
cow gamma globulins, cow albumin, hydrogen peroxide, dishes for
blot incubation, and gelatin. The PROCELL Electrophoresis Unit and
Electrophoresis Package 1/8 are needed but not provided.
CAT. NO.
DESCRIPTION
IND-4 Development of the Immune System
PRICE
$103.99
Hydrogen Peroxide
Dishes for Blot Incubation
Gelatin
Exp-802
Sample Buffer
Chicken Egg White
Quail Egg White
Turkey Egg White
Chicken Serum
Chicken Ovalbumin
Chicken Conalbumin
Antibodies to Ovalbumin
HRP Coupled Antibodies
Nitrocellulose
Blotting Paper
Protein Blot Stain
Tris Buffer Saline (TBS)
TBS + NP40
Color Development Buffer
Chloronapthol
Hydrogen Peroxide
Dishes for Blot Incubation
Gelatin
Exp-803
Calf Thymus
Calf Serum
Extraction Buffer
(2 Tubes)
Sample Buffer
LDH – Substrate
Calf Heart LDH
Calf Muscle LDH
EXP 804
Horse Radish
Peroxidase – Basic
Horse Radish
Peroxidase –
Mixture
Sample Buffer
Extraction Buffer
(2 Tubes)
Corn Seeds
Hydrogen Peroxide
4M Tris
Chloronapthol
Cytochome C
Hemoglobin-Serum
Albumin
For fast dependable service:
Call toll free: 800-733-6544
FAX: 765-523-3397
Visit: www.modernbio.com
&BTU4PVUIt-BGBZFUUF*/
IND5. SPECIFICITY OF ALBUMIN BINDING.
The binding of an enzyme to its substrate is only one example of
the many specific molecular interactions that occur in biological
systems. An analogous binding process occurs with serum
albumin which binds certain small molecular weight compounds
and serves as a carrier molecule for these compounds in blood.
In this exercise, students use an electrophoretic assay to examine
the binding of various dyes to albumin. The results of this graphic
analysis show that the binding of dyes to albumin is saturable,
specific, compatible, and dependent on the native structure of
the protein. The exercise is designed such that each of the eight
groups of students performs a different experiment. Each group
then describes their results and conclusion to the entire class.
This exercise is a valuable experience in analyzing data and
provides a fine introduction to enzyme kinetics. The analysis can
be performed during a single 3-hour or two 2-hour laboratory
session and includes: bovine serum albumin, dilution buffer,
bromophenol blue, orange G, ponceau S, sodium dodecyl sulfate,
and rabbit hemoglobin. Electrophoresis Package 1/8 is needed
but not provided.
CAT. NO.
DESCRIPTION
IND-5 Specificity of Albumin Binding
s&!8s7EBSITEWWWMODERNBIOCOMs
PRICE
$62.39
PROTIENS
Each of the individual experiments is supplied with the chemicals
and laboratory guides needed for 16 students working in pairs. If
you chose one or more of the experiments below, you should also
order Electrophoresis Package 1/8. Electrophoresis Package 1/8
provides sufficient agarose, gel stain and electrophoresis buffers
for up to 6 of the individual experiments in this series and in
Standard Laboratory Program 1.
Ep-1/8
Agarose
Gel Stain (Coomassie Blue)
Electrophoresis Buffer
Exp- 801
Goat Serum
Sheep Serum
Cow Serum
Horse Serum
Chicken Serum
Cow Transferrin
Cow Gamma Globulins
Cow Albumin
Antibodies to Cow
Albumin – HRP
Anti-Cow Gamma
Globulins – HRP
Nitrocellulose
Blotting Paper
Protein Blot Stain
Tris Buffer Saline (TBS)
TBS + NP40
Color Development Buffer
Chloronapthol
14
Molecular Biology of Proteins
Standard Laboratory Program 2
PRO
ELECTR TEIN
OP
-DENA HORESIS
TURIN
G-
A common procedure used for determining the size of proteins in the research laboratory is polyacrylamide gel electrophoresis,
where denatured proteins are separated in the presence of the detergent sodium dodecyl sulfate (SDS). However, preparing
polyacrylamide gels is laborious and requires a number of toxic chemicals. To avoid these limitations, we provide a special
blend of nontoxic agarose that yields excellent resolution of SDS-treated proteins. In this innovative series, students use SDSagarose gel electrophoresis to perform procedures including molecular weight determinations, peptide mapping analysis,
detection of specific enzymes in crude cell extracts, affinity chromatography, and immunological studies using the Western
blot procedure.
PROTIENS
201. Molecular Weight Determination
EXPERIMENTS
A first step in characterizing a protein often involves determining
its molecular weight. From this information, different proteins
may be compared and the number of amino acid residues in
a protein can be determined. Here, students determine the
molecular weight of two unknown proteins by comparing
their electrophoretic migration with the migration of standard
proteins. The protein standards and unknowns have been prestained so that your students can follow their progress during the
separation as shown below.
1 2 34 567 8
1 2 34 567 8
5
25
1 2 34 567 8
45
1 2 34 567 8
65
ELECTROPHORESIS TIME (MINUTES)
Prestained protein standards of known molecular weights and two proteins of
unknown molecular weights were subjected to electrophoresis for the indicated
times. The molecular weights of the unknown proteins can be determined by
comparing their migration with the migration of the standard proteins.
202. Identifying Sex-Specific Proteins
Vitellogenin is a protein produced in hen liver under the influence
of estrogen. This sex-specific protein enters the circulatory system
and is transported to the ovaries where it is broken down into
the egg yolk proteins lipovitellin and phosvitin. In this exercise,
students compare the proteins in egg yolk and in hen and rooster
sera by electrophoresis and identify vitellogenin, lipovitellin and
phosvitin. This exercise illustrates the use of electrophoresis
for protein identification and introduces concepts in molecular
endocrinology.
203. Comparing Human and Bacterial Amylase
Amylase in animals, plants and bacteria can be detected and
characterized by the procedure described in this exercise. The
analysis is performed by incorporating starch into an SDS-agarose
gel prior to electrophoresis of the denatured proteins. After
electrophoresis, the SDS is removed and the enzyme spontaneously
renatures. The location of amylase in the gel is then carried out
by staining the starch in the gel with iodine. Zones of enzyme
activity are devoid of starch and are seen as clear bands against a
background of blue. Using this procedure, students determine the
molecular weight of amylase in their own saliva and compare this
value to the molecular weight of amylases produced by bacteria
204. Peptide Mapping Analysis
SDS gel electrophoresis is used extensively to separate and
identify denatured proteins. However, because this method relies
on protein size alone, little information about proteins with the
same molecular weight can be obtained. Peptide mapping is
one of a number of techniques used to study the relatedness of
similarly sized proteins. With this method, proteases are used to
1
2 3 4 5 6 7 8
9 10 11 12 13 14 15 16
PEPTIDE MAPPING ANALYSIS.
Serum Albumin from human (lanes 2-4), cow (lanes 6-8), chicken (lanes 1012), and pigeon (lanes 14-16) were digested with chymotrypsin prior to this
separation by electrophoresis. Standard proteins are in lanes 1, 5, 9, and 13.
s-ODERN"IOLOGY)NCss
15
cut proteins into smaller peptide fragments and the fragments
derived from two or more proteins are compared. The number
and size of fragments generated from a protein are determined
largely by the protein’s amino acid sequence, since proteases
break peptide bonds adjacent to preferred amino acid residues.
In this exercise, students use peptide mapping to compare the
structural relatedness of serum albumin from the human, cow,
chicken and pigeon. Typical results of this exercise are shown
in the figure at the bottom of the previous page.
We now sell extra nitrocellulouse
for experiment 205.
See page 62 for details.
about two 2-3 hour
laboratory periods.
206. Affinity Chromatography
Purified proteins are often needed in the basic research laboratory
and for diagnostic and therapeutic procedures. An effective
technique for protein purification is affinity chromatography,
which exploits a specific interaction between a protein and a
complementary binding molecule. In this exercise, students
isolate albumin from horse serum by affinity chromatography
using a column matrix containing a reactive blue dye which binds
specifically to the albumin molecule. They then use electrophoresis
to analyze the isolated protein in order to verify the effectiveness
of the procedure.
This exercise requires approximately
two 2-3 hour laboratory periods.
EP-2
Boiling Rack
Agarose (nusive)
Gel Stain (Coomassie blue)
Electrophoresis Buffer
Gel Buffer
Push Pins
Exp 201
Unknown Protein #1
Unknown Protein #2
Standard Proteins
Rabbit Serum
EXP 202
Hen Serum Proteins
Rooster Serum Proteins
Egg Yoke Sample
Standard Proteins
Sample Buffer
EXP 203
Starch Solution
Potassium Iodine
Bacterial Amylase
Standard Proteins
Sample Buffer
Gel Incubation Buffer (2 Tubes)
EXP 204
Bovine Serum Albumin
Human Serum Albumin
Chicken Serum Albumin
Pigeon Serum Albumin
Chymotrypsin
Standard Proteins
Sample Buffer
EXP 205
Cow Serum
Horse Serum
Sheep Serum
Goat Serum
Donkey Serum
Antibodies to Cow
Albumin – HRP
Chicken Serum
Protein Blot Stain
Dishes for Blot Incubation
Gelatin
Tris Buffer Saline (TBS)
TBS + NP40
Chloronapthol
Hydrogen Peroxide
Color Development Buffer
Standard Proteins
Nitrocellulose
Blotting paper
Exp 206
Affi-gel Blue
Chromatographic Columns
Elution Buffer Concentrate
Standard Proteins
Sample Buffer
Horse Serum
Column Buffer
Price List - Standard Laboratory Program 2
CAT. NO.
DESCRIPTION
PRICE
S2
The Chemical Package for 8 pairs of
$243.58
students plus one student manual
and one instructor manual.
S2-BK 8 Student Manuals
$63.14
S2-SM Sample Student Manual plus one instructor manual. $9.61
INDIVIDUAL EXPERIMENTS
Each of the individual experiments is supplied with the chemicals
and laboratory guides needed for 16 students working in pairs. If
you chose one or more of the experiments below, you should also
order Electrophoresis Package 2M or Electrophoresis Package
2. Electrophoresis Package 2M provides sufficient agarose,
gel stain, and electrophoresis buffers for 1 of the individual
experiments in this series (four gels with 15ml of agarose per
gel). Electrophoresis Package 2 provides sufficient agarose, gel
stain, and electrophoresis buffers for up to 6 of the individual
experiments in this series.
EP-2M
EP-2
EXP-201
EXP-202
EXP-203
EXP-204
EXP-205
EXP-206
Electrophoresis Package 2M
Electrophoresis Package 2
Molecular Weight Determination
Identifying Sex-Specific Proteins
Comparing Human and Bacterial Amylase
Peptide Mapping Analysis
Protein Evolution and the Western Blot
Affinity Chromatography
$48.57
$135.65
$54.86
$56.53
$55.45
$71.77
$94.56
$87.06
Note that some of these experiments can also be performed using
SDS-polyacrylamide gel electrophoresis. Please see page 16 for details.
s&!8s7EBSITEWWWMODERNBIOCOMs
PROTIENS
205. Protein Evolution and the Western Blot
The Western blotting procedure is rapidly replacing conventional
methods for identifying and characterizing specific proteins in
complex protein mixtures. This technique is used extensively
for this purpose in the research laboratory and is increasingly
being used in diagnostic medicine for detecting proteins of
disease agents such as the structural core proteins of the AIDS
virus. Here, students will perform this technique to examine the
evolutionary distance between different mammals. Guided by
step-by-step instructions, they subject serum proteins from six
species to gel electrophoresis and transfer the separated proteins
to nitrocellulose membranes. They then use an enzyme-linked
immunoassay to compare the extent to which the separated
albumins in the serum samples are related to those from cow. This
exercise provides an exciting lesson in molecular evolution and
introduces your students to one of the most important techniques
This exercise requires
of molecular biology.
Chemical Package Contents
16
Polyacrylamide Gel Electrophoresis
Protein Analysis
PROTIENS
In the experiments described in this series, denatured proteins are separated according to size on high-resolution SDS
polyacrylamide gels. Students perform procedures including molecular weight determinations, peptide mapping analysis,
isolation and characterization of specific cellular proteins and immunological studies using the Western blot procedure. Each
experiment was designed so that the protein samples of 8 groups of students are analyzed on two or four electrophoretic
gels. (purchase seperately through Bio-Rad, see the bottom of page 17.) Electrophoresis equipment and an electrophoresis
package containing electrophoresis buffer and gel stain can be purchased from pages 48-50 in this catalog.
201P. Molecular Weight Determination
A first step in characterizing 1 2 3 4 5 6 7 8 9 10 11 12
a protein often involves
determining its molecular
weight. From this information,
different proteins may be
compared and the number
of amino acid residues in a
protein can be determined.
Here, students determine
the molecular weight of two unknown proteins by comparing
their electrophoretic migration with the migration of standard
proteins. The protein standards and unknowns have been
pre-stained so that your students can follow their progress
during the separation. This experiment includes instructions,
prestained Unknown Protein #1,Unknown Protein #2, Standard
Proteins, and Rabbit Serum. Requires
Staining is not
three 12 well gels for 8 groups of
necessary.
students or two gels for 6 groups.
CAT. NO.
DESCRIPTION
EXP-201P Molecular Weight Determination
PRICE
$54.86
IND-11P. Contractile Proteins from Cow Heart
In this experiment, students prepare a protein extract from
cow heart. They then determine the molecular weights of major
contractile proteins by 1 2 3 4 5 6 7 8 9 10 11 12
comparing their migration
on SDS-polyacrylamide gels
to the migration of standard
proteins of known size.
Students also identify and
determine the molecular
weight of the major proteins
found in milk. This exercise
requires 1 three-hour or 2 two-hour lab sessions and a table
top centrifuge or microcentrifuge is required. Typical results
of the exercise are shown on the gel above. Includes cow
heart, milk, standard proteins, sample buffer, and instructions.
Requires three 12 well gels for 8 groups of students or two gels
for 6 groups.
CAT. NO.
DESCRIPTION
IND-11P Contractile Proteins from Cow Heart
PRICE
$72.10
IND-25P. Isolation of Chromosomal Proteins
The five histone molecules are the major proteins that associate with DNA to form nucleosomes. In this experiment,
students isolate nuclei from chicken erythrocytes and then
prepare nucleosomes from the erythrocytes by using a simple
procedure that employs micrococcal nuclease. They then determine the molecular weights of the five histone proteins by
1
2
3
4
5
6
7
8
9 10
11 12
comparing their migration on SDS-polyacrylamide gels to the
migration of standard proteins of known size. This exercise
requires a single two-three hour lab session and a microcentrifuge is required. The exercise provides a fine introduction to the nucleosome structure of chromatin and typical
results are shown on the gel above. This experiment includes
chicken erythrocytes, standard proteins, sample buffer, nuclear isolation buffer, micrococcal nuclease and instructions.
Requires two 12 well gels for 8 groups of students.
CAT. NO.
DESCRIPTION
IND-25P Isolation of Chromosomal Proteins
PRICE
$55.18
Electrophoresis Package for SDS Polyacrylamide Gels
The package contains Tris-Glycine-SDS electrophoresis buffer
(makes 3 liters) and Coomassie Blue Gel Stain (makes 1 liter).
The package provides sufficient materials for three of our
polyacrylamide experiments. The gels must be ordered separately.
CAT. NO.
DESCRIPTION
EP-2P Electrophoresis Package for
SDS Polyacrylamide Gels
s-ODERN"IOLOGY)NCss
PRICE
$29.41
17
CAT. NO.
DESCRIPTION
EXP-204P Peptide Mapping Analysis
PRICE
$71.77
205P. Protein Evolution and the Western Blot
The Western blotting procedure is rapidly replacing conventional
methods for identifying and characterizing specific proteins in
complex protein mixtures. This technique is used extensively for
this purpose in the research laboratory and is increasingly being
used in diagnostic medicine for detecting proteins of disease
agents. Here, students will perform this technique to examine
the evolutionary distance between different mammals. Guided by
step-by-step instructions, they subject serum proteins from six
species to gel electrophoresis and transfer the separated proteins
to nitrocellulose membranes. They then use an enzyme-linked
immunoassay to compare the extent to which the separated
albumins in the serum samples are related to those from cow. This
exercise provides an exciting lesson in molecular evolution and
introduces your students to one of the most important techniques
of molecular biology. This exercise requires about two 2-3 hour
laboratory sessions. The exercise was designed for 8 groups of
students and includes: sera from cow, horse, sheep, goat, chicken
and donkey, standard proteins, anti-cow albumin (peroxidase
linked), nitrocellulose, blotting paper, protein blot stain, Tris
buffer saline (TBS), TBS + NP40, color development buffer,
chloronapthol, hydrogen peroxide, dishes for blot incubation,
and gelatin. Requires three 12 well gels for 8 groups of students.
CAT. NO.
DESCRIPTION
EXP-205P Protein Evolution and the Western Blot
PRICE
$96.30
206P. Affinity Chromatography
Purified proteins are often needed in the basic research laboratory
and for diagnostic and therapeutic procedures. An effective
technique for protein purification is affinity chromatography,
which exploits a specific interaction between a protein and a
complementary binding molecule. In this exercise, students
isolate albumin from horse serum by affinity chromatography
using a column matrix containing a reactive blue dye, which
binds specifically to the albumin molecule. They then use
electrophoresis to analyze the isolated protein in order to verify
the effectiveness of the procedure. This experiment includes Affigel Blue, chromatographic columns, column buffer, elution buffer,
standard proteins, horse serum and instructions. Requires three
12 well gels for 8 groups of students or two gels for 6 groups.
CAT. NO.
DESCRIPTION
EXP-206P Affinity Chromatography
PRICE
$87.06
IND-27 Composition of the Nuclesome
The primary level of chromosome structure in eukaryotes
occurs when the DNA molecule 1 2 3 4 5 6 7 8 9 10 11 12
is wrapped around histone
proteins into particles called
nucleosomes. Evidence for
this “beads on a string” model
is derived from nuclease
digestion studies. When nuclei
are incubated with micrococcal
nuclease, the enzyme cleaves
the linker DNA between
nucleosomes (the string) but not the nucleosomal core DNA (the
beads). With this background information, students isolate nuclei
from chicken erythrocytes and then prepare nucleosomes from
the nuclei by using a simple procedure that employs micrococcal
nuclease. They then analyze the resulting nucleosome DNA
fragments by agarose gel electrophoresis. They also determine
the molecular weights of the five histone proteins associated
with the isolated nucleosomes by comparing their migration on
SDS-polyacrylamide gels to the migration of prestained standard
proteins of known size. Sufficient chemicals are provided so that
this experiment can be performed twice by 8 groups of students.
This exercise requires a microcentrifuge, EP-2P, EP-3/4 and
three 12-well polyacryamide gels. This exercise provides a fine
introduction to structure and composition of the nucleosome
and typical results are shown on both the DNA and protein gels.
CAT. NO.
DESCRIPTION
PRICE
IND-27 Comosition of the Nuclesome
$56.36
Includes detailed instructions, chicken erythrocytes, prestained
standard proteins, protein sample buffer, DNA sample buffer,
calf thymus nucleosome standard DNA, nuclear isolation buffer,
micrococcal nuclease, nuclear stain, EDTA, small transfer pipets
and large transfer pipets.
Precast Polyacrylamide gels can be ordered from
Bio-Rad. (800-424-6723) Their catalog number is 1611177.
s&!8s7EBSITEWWWMODERNBIOCOMs
PROTIENS
204P. Peptide Mapping Analysis
SDS gel electrophoresis is used extensively to separate and identify
denatured proteins. However, because this method relies on
protein size alone, little information about proteins with the same
molecular weight can be obtained. Peptide mapping is one of a
number of techniques used to study the relatedness of similarly
sized proteins. With this method, proteases are used to cut
proteins into smaller peptide fragments and the fragments derived
from two or more proteins are compared. The number and size
of fragments generated from a protein are determined largely
by the protein’s amino acid sequence, since proteases break
peptide bonds adjacent to preferred amino acid residues. In this
exercise, students use peptide mapping to compare the structural
relatedness of serum albumin from the human, cow, chicken
and pigeon. Typical results of the exercise are shown below. This
experiment includes human, cow, chicken and pigeon albumins,
1 2 3 4 5 6 7 8
standard proteins, 1 2 3 4 5 6 7 8
sample buffer and
chymotrypsin.
Requires three
12 well gels for 8
groups of students
or two gels for 6
groups.
18
Molecular Biology of Nucleic Acids
Standard Laboratory Program 3
This outstanding program provides a solid foundation in the molecular biology of DNA. Your students will investigate
such contemporary topics as restriction nuclease mapping, the complexity of the prokaryotic and eukaryotic genomes
and the nucleosome structure of the chromosome. The studies on plasmid amplification using recombinant DNA
techniques enable students to conduct the same procedures that form the basis of the biotechnology industry.
EXPERIMENTS
301. The Length of DNA Molecules
Electrophoresis in agarose gels is the most common method used
for determining the size of DNA molecules. In this introductory
exercise, students determine the length of an unknown DNA molecule
by comparing its electrophoretic mobility with six DNA molecules of
known size as shown below.
Length of DNA EXP-301
1 2 3 4 5 6 7 8
Restriction Nuclease Mapping EXP-302
1 2 3 4 5
6 7 8
303. Plasmid DNA Structure
Plasmids are small circular DNA molecules found in most bacteria. A
plasmid can exist in different structural states and these states can be
distinguished by their migration on agarose gels. In this experiment,
students study these structures using enzymes and electrophoresis and
show that the structures are interconvertible. This exercise provides
an introduction to higher-order DNA structure which is thought to be
important in the control of transcription and replication in bacteria
and, perhaps, in higher organisms as well.
New Plasmid
DNA
304. Molecular Cloning
DNA fragments of known lengths
(lanes 1, 3, 5, and 7) and unknown
length (lanes 2, 4, 6, 8) are separated
by electrophoresis.
Lambda phage DNA was digested with
EcoR1 or EcoR1 plus BamH1 prior to this
electrophoretic separation.
DNA STAINING PROCEDURE
Our programs use a safe and simple two-step procedure for
staining DNA in agarose gels. First, a nontoxic purple dye is
incorporated into the agarose gel prior to electrophoresis.
Faint purple DNA bands can be clearly seen after 30 minutes
of electrophoresis. This rapid procedure enables the student to
obtain preliminary results of the experiment in a short period
of time. After the electrophoretic run, gels are incubated with
methylene blue to complete the staining procedure.
Isolating Procedure.
This two part exercise provides state-of-the-art information and
practical experience with a variety of techniques that form the
foundation of the biotechnology industry. In part A, students create a
strain of E.coli that is resistant to the antibiotic ampicillin by introducing
a plasmid that contains an ampicillin-resistance gene. The success
of the transformation is monitored by growing the bacteria on an
ampicillin-containing media. This experiment provides sufficient sterile
materials for sixteen platings. In Part B, students isolate the amplified
plasmid from the bacteria without the use of toxic chemicals. Then they
digest the isolated DNA with EcoR1 and compare the electrophoretic
properties of linear and circular plasmid DNA molecules. By inserting
desired DNA segments into plasmids, this procedure has enabled
scientists to amplify more than 1000 specific genes including those for
human interferon, insulin, and growth hormone. Part B of the exercise
requires a centrifuge that can be operated at a force of at least 3,000
x g (such as a microcentrifuge or a larger floor model). The entire
exercise requires three 2-3 hour laboratory periods and the materials
provided are shown below.
Electrophoresis
MATERIALS FOR EXP 304
302. Restriction Nuclease Mapping of DNA
Bacteriophage lambda is a DNA virus that attacks E.coli. Here, students
dissect lambda DNA using the restriction endonucleases EcoR1 and
BamH1 in order to identify specific sites, sequences, and structures along
the phage genome. This single exercise enables students to explore a
number of exciting topics in molecular biology, including the specificity
of restriction endonucleases, DNA mapping strategies, complementary
base-pairing of DNA, and the structure of a viral genome.
s-ODERN"IOLOGY)NCss
equipment not
required.
19
305. Identifying Satellite Sequences
A large fraction of the DNA of vertebrates consists of nucleotide
sequences that are repeated thousands or more times in the
genome. Satellite sequences are a major class of these repeated
elements and these sequences are found in most eukaryotic
organisms. There are about 1 million cutting sites for the
restriction enzyme EcoRI in the typical vertebrate genome
and about 1 million DNA fragments are generated following
digestion of vertebrate DNA with this enzyme. When this DNA is
separated by electrophoresis, the large number of differentlysized fragments appear as a background smear on the gel lane.
However, as shown in the figure to the right, DNA fragments
from the highly repeated satellites form discrete bands because
they are the same size. In this exercise, students identify satellite
sequences in the cow genome and determine if such sequences
are present in chicken DNA.
EP-3/4
Agarose
Gel Stain (Methylene Blue)
In Gel Stain
Electrophoresis Buffer
Exp 301
Dye Mixture
DNA Markers
Unknown DNA
Exp 302
Phage Lambda DNA
Bam H1
Sample Buffer
EcoR1
Restriction Nuclease Buffer
Exp 303
Plasmid DNA
Sample Buffer
S1 Nuclease
Dye Mixture
EcoR1
Restriction Nuclease Buffer
S1 Nuclease Buffer
Exp 304 Part A
E.coli
Plasmid pUC18
Exp 304 Part B
Ampicillin
E.coli -pUC18
Calcium Chloride
Nutrient Broth +
Nutrient Broth
Ampicillin
Perti Dishes
Electrophoresis Sample
Nutrient Agar + Ampicillin
Buffer
Inoculating Loops
Dye Mixture
Sterile Transfer Pipets
Restriction Nuclease
Sterile Tubes
Buffer
Exp 305
EcoR1
Calf DNA
Isopropanol
Chicken DNA
Plasmid Isolation
EcoR1
Restriction Nuclease Buffer (found in green box)
Solution I - 1 bottle
Dye Mixture
Solution II - 1 bottle
Sample Buffer
Solution III - 1 bottle
Exp 306
Equilibration Buffer - 1
Nucleosome DNA # 1
bottle
Nucleosome DNA # 2
Buffer HB - 1 bottle
Nucleosome DNA # 3
Nuclear Incubation Buffer (2) Wash Buffer - 1 bottle
Elution Buffer - 1 bottle
Dye Mixture
HiBind DNA Column - 24
Sample Buffer
Collection Tubes - 24
Wheat Germ
RNase - 1 Tube
Micrococcal Nuclease
Cheese Cloth
Transfer Pipets
Nucleosome DNAEXP-306
1 2 3
4 5 6 7 8
DNA from calf thymus (even numbered Note the ladder pattern produced when the
lanes) was digested with EcoR1 prior DNA in chromatin is cleaved by micrococcal
to this electrophoretic separation. nucleases. (lanes 2-4, 6-8).
Note the discrete satellite DNA band.
Price List - Standard Laboratory Program 3
CAT. NO.
DESCRIPTION
S3
The Chemical Package for 16 students
working in pairs plus one student manual and
one instructor manual (The student manual
may be reproduced for educational purposes.)
S3-BK 8 Student Manuals
S3-SM One Student Manual plus one instructor manual.
PRICE
$239.35
$63.14
$9.61
INDIVIDUAL EXPERIMENTS
Each of the individual experiments is supplied with the chemicals
and laboratory guides needed for 16 students working in pairs.
(Please see page 21 for additional details). If you chose one or more
of the experiments below, you should also order Electrophoresis
Package 3/4. Electrophoresis Package 3/4 provides sufficient
agarose, gel stains and electrophoresis buffers for up to 6 of the
individual experiments in this series. The package also includes
the dye for rapid DNA staining during electrophoresis (See
facing page). Electrophoresis Package 3/4 is also suitable for the
experiments in Standard Laboratory Programs 4 and 10 and the
experiments on pages 20-22.
EP-3/4 Electrophoresis Package 3/4
EXP-301 The Length of DNA Molecules
EXP-302 Restriction Nuclease Mapping of DNA
EXP-303 Plasmid DNA Structure
EXP-304 Molecular Cloning (Part A + Part B)
EXP-304A Molecular Cloning(Part A only)
EXP-304B Molecular Cloning(Part B only)
EXP-305 Identifying Satellite Sequences
EXP-306 The Nucleosome Structure of Chromatin
s&!8s7EBSITEWWWMODERNBIOCOMs
$67.07
$48.97
$62.48
$62.48
$99.86
$71.18
$62.24
$56.37
$71.77
DNA
306. The Nucleosome Structure of Chromatin
The primary level of chromosome structure in eukaryotes occurs
when the DNA molecule is wrapped around histone proteins
into particles called nucleosomes. Evidence for this "beads on a
string" model is derived from nuclease digestion studies. When
nuclei are incubated with micrococcal nuclease, the enzyme
cleaves the linker DNA between nucleosomes (the string) but not
the nucleosomal core DNA (the beads). When the DNA isolated
from these nuclei is electrophoresed on agarose gels, the DNA
exhibits a ladder pattern with the length of the DNA in each band
representing a multiple of the nucleosomal unit as shown in the
Figure on this page. With this background information, students
first examine nucleosomal DNA from calf thymus that is given
in this chemical package. They then prepare nuclei from wheat
germ, incubate them with micrococcal nuclease and examine
the ladder pattern in the DNA by electrophoresis. This exercise
requires two 2 - 3 hour laboratory periods.
Chemical Package Contents
Satellite DNA EXP-305
1 2 3 4 5 6 7 8
20
The New Genetics
Standard Laboratory Program 10
The exercises in this program are designed to illustrate key principles of molecular genetics including genome
organization, gene function, and DNA fingerprinting. Each exercise emphasizes an investigative approach where students
draw conclusions from experimental results. Each exercise also illustrates fundamental biological concepts. The four
exercises can be done independently or in sequence and are recommended for beginning and intermediate-level students.
DNA
1001. Anatomy and Evolution of the Genome
Common plasmids are simple DNA molecules which contain a
few genes and regulatory elements. Most viral genomes are more
complex. For example, the genome of phage lambda contains
approximately 50 genes. About 4,000 genes are present in the
E.coli genome while there is approximately 1,000 times more
DNA in the genome of a mammal. This progression in genome
complexity is the topic of this exercise. Here, students compare the
electrophoretic patterns of restriction digests of a plasmid, phage
lambda DNA, and cow DNA from thymus and kidney as shown in
the figure below. The exercise serves as a good introduction for
determining the size of DNA molecules and provides an appreciation
for the complexity of genomes from different organisms.
1002. Analysis of a Genome Segment
Phage lambda DNA and a recombinant plasmid containing a segment
of the phage lambda genome are provided as the starting point for
this exercise. Students digest these DNAs with EcoR1 and BamH1
and then analyze the fragments as shown below. Determination of
the size of the fragments enables the student to identify the precise
region in the lambda genome that is contained within the plasmid.
Anatomy and Evolution of the
(FOPNFt&91
1 2 3 4 5 6 7 8
Analysis of a Genome Segment
EXP-1002
1 2 3 4 5 6 7 8
1003. DNA Fingerprinting (An authentic analysis)
DNA fingerprinting relies on the fact that DNA can be extremely
variable in sequence from one individual to another and from one
organism to another. Thus, the method has been used to identify
individuals in forensic and paternity cases and to study genetic
lineages of closely related organisms. These applications are
discussed in the text and the latter application is illustrated in this
experiment. In the exercise, students are given two DNA samples
which are identified in the instructor guide. One sample is from
sheep and the other from cow. In part A, the DNAs are digested with
restriction enzymes and the sizes of the bands composed of highly
repeated sequences are determined by comparison to standard
DNAs of known lengths as shown below. In part B, the nonrepeated
sequences are eliminated by a simple novel procedure. This step
serves to enhance the banding patterns of the repeated sequences
as shown in the gel on the right. From the results of this analysis,
students determine which DNA sample is from cow and which
is from sheep. This exercise requiring two 2-3 hour laboratory
periods is the only authentic (non-simulated) DNA fingerprinting
exercise available for the
See page 62 to make up your own DNA
fingerprinting story with experiment 3-20.
teaching laboratory.
%/"'JOHFSQSJOUJOHt&91
Part A
Part B
1 2 3 4 5 6 7 8
DNA from plasmid pUC18 (lanes
1 & 5), lambda phage (lanes 2
& 6). calf thymus (lanes 3 & 7),
and calf kidney (lanes 4 & 8) are
separated by electrophoresis. The
DNA samples for this exercise are
supplied ready for electrophoresis.
Lambda phage DNA (lanes 1, 2,
5, & 6) or a plasmid containing a
segment of the phage genome
(lanes 3, 4, 7, & 8) were digested
with EcoR1 or EcoR1 + BamH1 prior
to this electrophoretic separation.
DNA from cow (lanes 3 & 4) and sheep
(lanes 6 & 7) were digested with EcoR1
or BamH1 prior to this electrophoretic
analysis. DNA markers are shown in
lanes 2 & 5.
s-ODERN"IOLOGY)NCss
1 2 3 4 5 6 7 8
Cow and sheep DNA are subjected to
a simple novel procedure in order to
eliminate nonrepeated sequences.
The repeated sequences were then
electrophoresed on this gel.
21
1004. Genotype to Phenotype
Watch a video about this popular
This exercise was designed to
transformation experiment at
MODERNBIO.COM
provide an exciting introduction
to specific gene structure and function. In the experiment,
students are given two plasmids (A and B) which are identified
in the instructors guide. One plasmid (A) has a functional gene
for the enzyme ß-galactosidase. The ß-galactosidase gene in the
other plasmid (B) is inactive because it contains a segment of
foreign DNA. In the first part of the exercise, students analyze
restriction digests of both plasmids in order to determine which
plasmid should have a functional ß-galactosidase gene. In the
second part of the exercise, the plasmids are introduced into
E.coli by transformation and the color of the resulting colonies
(blue or white) is then used to assess the functional status of the
ß-galactosidase gene. This exercise can be completed within a
single 3-hour laboratory session or two 2-hour laboratory periods.
Price List - Standard Laboratory Program 10
CAT. NO.
DESCRIPTION
PRICE
S10
The Chemical Package for 16 students
$232.04
working in pairs plus one student manual and one
instructor manual (The student manual may be
reproduced for educational purposes).
S10-BK 8 Student Manuals
$63.14
S10-SM Sample Student Manual plus one instructor manual. $9.61
For fast dependable service:
Call toll free: 800-733-6544
FAX: 765-523-3397
Visit: www.modernbio.com
&BTU4PVUIt-BGBZFUUF*/
INDIVIDUAL EXPERIMENTS
Each of the individual experiments are supplied with the
chemicals and laboratory guides needed for 16 students
working in pairs. If you chose one or more of the experiments
below, you should also order Electrophoresis Package 3/4. EP3/4 provides sufficient agarose, gel stain and electrophoresis
buffers for up to 6 of the individual experiments in this
series. Electrophoresis Package 3/4 is also suitable for the
experiments in Standard Laboratory Programs 3 and 4.
EP-3/4
$67.07
EXP-1001 Anatomy and Evolution of the Genome
$64.91
EXP-1002 Analysis of a Genome Segment
$73.37
EXP-1003 DNA Fingerprinting (An authentic analysis)
$82.46
EXP-1004 Genotype to Phenotype
$87.60
Chemical Package Contents
White
Colonies
Blue
Colonies
EP-3/4
Agarose
Gel Stain (Methylene Blue)
In Gel Stain
Electrophoresis Buffer
EXP-1001
Plasmid DNA-EcoR1 Cut
Phage Lambda DNA-Hind III Cut
Calf Thymus DNA-EcoR1 Cut
Calf Kidney DNA-EcoR1 Cut
EXP-1002
Lambda DNA
Plasmid-Lambda
EcoR1
BamH1
Restriction Nuclease Buffer
Sample Buffer
EXP-1003
Unknown DNA 1
Unknown DNA 2
EcoR1 Endonuclease
BamH1 Endonuclease
Restriction Endonuclease
Buffer (2)
s&!8s7EBSITEWWWMODERNBIOCOMs
Sample Buffer
S1-Nuclease
S1-Nuclease Buffer
DNA Markers
EXP-1004
Plasmid A
Plasmid B
Plasmid C
EcoR1 Endonuclease
Endonuclease Buffer
Sample Buffer
E.coli
Ampicillin
DNA Marker
CaCl2
Nutrient Broth
Nutrient Agar + Ampicillin
X-gal
X-gal Solvent
Inoculating Loops
Sterile Transfer Pipets
Sterile Tubes
Petri Dishes
DNA
Electrophoresis Package 3/4
22
DNA Hybridization Analysis
Standard Laboratory Program 4
The revolution brought about by molecular biology depended heavily on nucleic acid hybridization procedures.
These techniques are used extensively in the research laboratory for detecting specific nucleotide sequences in
DNA and RNA and are increasingly being applied in medicine for diagnosing disease. Modern Biology, Inc. is proud
to introduce these methods into your teaching laboratory. In this laboratory program, students use hybridization
procedures to investigate key topics in contemporary biology including the evolution of the vertebrate genome, the
complexity of the viral chromosome, and the analysis of a specific nucleotide sequence in mammalian DNA. The
package comes complete with all of the biochemicals that you will need although a water bath incubator that will
maintain a temperature of 60-65°C is required for these experiments. The procedures used in these exercises are
safe for your students because the four hybridization probes provided with the chemical package were prepared
using a nonradioactive labeling system.
DNA
EXPERIMENTS
401. Evolution of the Vertebrate Genome
The history of evolution is recorded in the genomes of presentday organisms and enlightened guesses of evolutionary events
can be made by comparing DNA sequences in different species.
Evidence drawn from this approach has led to the construction of
family trees of organisms that agree remarkably well with those
obtained from more traditional procedures. In fact, on a number
of occasions, comparative DNA sequence analysis has been used
to clarify and expand on phylogenetic relationships that were
derived from classical studies. For example, although biologists
have long disagreed about the taxonomic placement of the
giant panda, recent studies using DNA hybridization techniques
strongly suggest that this animal is more closely related to bears
than to raccoons. These important concepts are illustrated in this
exercise where students use a dot-blot hybridization procedure to
compare DNA sequences in salmon, turkey, chicken and cow. In
the analysis, single strands of chicken or cow DNA that have been
previously linked to biotin are mixed with single strands of DNA
from a second species. The two types of DNA are allowed to form
double-stranded hybrid molecules and the extent of hybridization
is measured using a simple color-producing enzyme reaction.
This exercise requires approximately two 2-3 hour laboratory
periods and electrophoresis equipment is not required.
402. Application of the Southern Blot Procedure
In l975, Edward M. Southern at the University of Edinburgh,
developed a powerful technique for DNA analysis which has
become known as Southern blotting. Here your students use
the Southern blotting procedure to identify the major control
region for transcription and replication in the lambda phage
Agarose Gel Southern Blot
Detection of the Control Region in Lambda Phage DNA
Left: Restriction nuclease digests of lambda DNA were
electrophoresed on an agarose gel and the gel was stained. Right:
The separated DNA fragments on the gel were transferred to a
nylon membrane and the DNA containing the control region was
identified by hybridization analysis.
genome. Following the step-by-step procedures in their manuals,
students digest lambda DNA with a restriction endonuclease,
electrophorese the DNA, and then transfer the separated fragments
on the gel to a nylon membrane. The DNA fragment containing the
control region is then identified by hybridization analysis using a
biotinylated probe for the control region and the enzyme-colorproducing assay. This exercise provides a wealth of practical
information on one of the most powerful methods in molecular
biology and illustrates an important strategy for mapping simple
and complex genomes. This experiment requires about three 2-3
hour laboratory periods and typical results are shown above.
s-ODERN"IOLOGY)NCss
23
403. Detecting a Specific Sequence in the Mammalian
Genome
Southern hybridization analysis can now be used for tracing defective
genes in human DNA including those involved in Huntington’s
disease, Duchenne Muscular Dystrophy, and Cystic Fibrosis. This
application is becoming increasingly important for diagnosing these
diseases in members of afflicted families and it seems likely that the
method will be used to detect additional abnormal and even normal
human genetic traits in the near future. In this exercise students
perform Southern hybridization analysis in order to characterize a
specific sequence in mammalian DNA. Students digest cow, sheep,
and chicken DNA with EcoRI, electrophorese the DNA, and transfer
the separated fragments to a nylon membrane. The DNA fragments
containing the cow satellite is then detected by hybridization using
a biotin-labeled probe made from a plasmid that contains the cow
satellite sequence. This exercise requires about three 2-3 hour
laboratory periods.
We now sell extra nylon membrane
and blotting paper (folder).
CAT. NO.
S4-BK
S4-SM
PRICE
Price List - Standard Laboratory Program 4
The Chemical Package for 16 students
working in pairs plus one student manual
and one instructor manual (The student manual
may be reproduced for educational purposes.)
8 Student Manuals
1 Student Manual and 1 Instructor Manual
$238.50
$63.14
$9.61
IND-16. Identifying Viral DNA by Rapid Southern Blotting
A limitation to the Southern blotting procedure for the teaching
laboratory is that it requires multiple laboratory sessions. The
procedures have been streamlined for this experiment so that they
can be carried out in a single 3-hour laboratory period. In the
exercise, students electrophorese three unknown DNA samples
on an agarose gel. One of the samples (instructor keyed) contains
biotinylated DNA from the bacterial virus lambda. Following
electrophoresis, the DNA in the gel is transferred to a nylon
membrane during a 15-minute blotting step and the viral DNA
on the blot is then detected by using an enzyme-color-producing
EP-3/4
Agarose
Gel Stain (Methylene Blue)
In Gel Stain
Electrophoresis Buffer
Exp 401
Cow DNA
Chicken DNA
Turkey DNA
Salmon DNA
Biotinylated Chicken DNA
Biotinylated Cow DNA
Hybridization Buffer
Tris Buffer
NP40
Hydrogen Peroxide
Gelatin
Chloronapthol
Hybridization Bags
Nylon Membranes
Blotting Paper
Avidin-Peroxidase
Methylene Blue
Paper Clips
Foam Rack
Exp 402
EcoR1 Endonuclease
Hybridization Buffer
Tris Buffer
NP40
Hydrogen Peroxide
Gelatin
Chloronapthol
Hybridization Bags
Nylon Membranes
Blotting Paper
Avidin-Peroxidase
4 Blotting Devices
Foam Rack
Paper Clip
Bam H1
Restruction Buffer
Electropharesis
Sample Buffer
Bioimulated Probe
Exp 403
Cow DNA
Chicken DNA (EcoR1 cut)
Sheep DNA (BamH1 cut)
EcoR1
EcoR1 Buffer
Sample Buffer
Biotinylated Satellite Plasmid
Hybridization Buffer
4 Blotting Devices
DNA Markers
Tris Buffer
NP40
Hydrogen Peroxide
Gelatin
Chloronapthol
Hybridization Bags
Nylon Membranes
Blotting Paper
Avidin-Peroxidase
Paper Clips
Foam Rack
INDIVIDUAL EXPERIMENTS
Each of the individual experiments is supplied with the chemicals
and laboratory guides needed for 16 students working in pairs.
If you chose one or more of the experiments on this page, you
should also order Electrophoresis Package 3/4. Electrophoresis
Package 3/4 provides sufficient agarose, gel stain and
electrophoresis buffers for up to 6 of the individual experiments
in this series. Note that Electrophoresis Package 3/4 is also
suitable for the experiments in Standard Programs 3 and 10.
EP-3/4
Electrophoresis Package 3/4
(for up to 6 individual experiments)
EXP-401 Evolution of the Vertebrate Genome
$67.07
$126.59
EXP-402 Application of the Southern Blot Procedure
$149.78
EXP-403 Detecting a Specific Sequence in the
Mammalian Genome
$140.87
reaction. The experiment was designed for 8 groups of students
and sufficient materials are provided so that four blots can be
prepared and analyzed. The exercise requires Electrophoresis
Package 3/4 and a water bath maintained at 37° C.
CAT. NO.
DESCRIPTION
IND-16 Identifying Viral DNA by Rapid Southern Blotting
PRICE
$119.19
Includes instructions, DNA samples 1-3, dye mixture, blotting
paper, nylon membranes, petri dishes,methylene blue, Tris-buffer
saline (TBS), TBS+NP-40, streptavidin peroxidase,
Hydrogen peroxide, chloronapthol, gelatin and Tris buffer.
s&!8s7EBSITEWWWMODERNBIOCOMs
DNA
S4
DESCRIPTION
Chemical Package Contents
24
Genetic Engineering
(Standard Laboratory Program 5)
Recombinant DNA techniques played a central role in the recent emergence of biology into the golden age. This
program enables the advanced student to approach these frontiers in biotechnology. The program provides a
challenging series of seven 3-hour laboratory sessions which are intended to give students hands-on-experience and
a detailed understanding of these new investigative techniques and their potentials.
In the program, students prepare a library of recombinant plasmids which contain different regions of the lambda
phage genome. They then amplify and characterize the lambda DNA in selected plasmids in order to identify the phage
genes that they contain. The basic strategy followed in the program is shown below and is one that has been used
by investigators around the world to clone DNA molecules from literally thousands of prokaryotic and eukaryotic
organisms. However, many of the procedures have been modified to make them safe and suitable for a college
laboratory class. The program provides essentially all of the instructions, chemicals, sterile media, and expendable
accessories that are needed to carry out DNA electrophoresis, restriction nuclease digestions, DNA ligations, bacterial
transformations, bacterial selection for ampicillin resistance and ß-galactosidase production, plasmid isolation, DNA
hybridization, and Southern blot procedures. A centrifuge that can be operated at a force of at least 3,000 x g (such
as a microcentrifuge or a larger floor model) and a water bath incubator that will maintain a temperature of 60-65°C
are required for the program. A shaking bacterial incubator is strongly recommended.
DNA
A typical laboratory schedule for this program is given
below where each lab session is approximately 3 hours.
Lab Session 1
Students digest phage lambda DNA with EcoR1 and ligate the
resulting DNA fragments to EcoR1-digested pUC18 in order to
create a series of recombinant plasmids. Some of the plasmids
in the mixture will lack inserted lambda DNA while the others
will have DNA inserts derived from various regions of the phage
genome.
Lab Session 2
The recombinant molecules are characterized by electrophoresis
and then introduced into competent E.coli cells by transformation.
The cells are plated onto nutrient agar containing ampicillin and X
gal. The ampicillin selects for all cells that have taken up plasmid
while the X gal permits students to identify colonies which contain
plasmids with lambda DNA inserts. The identification is based on a
powerful color selection assay. Plasmid pUC18 contains a portion
of the lac Z gene which codes for ß-galactosidase and X gal is a
substrate of this enzyme. When pUC18 is carried by appropriate
strains of E.coli, the lac Z gene is active and the ß-galactosidase
converts X gal to a blue product which gives rise to blue colonies.
If foreign DNA is inserted into the EcoR1 site of pUC18, the lac Z
gene is inactivated and the colonies remain white.
Lab Session 3
Each student prepares overnight cultures from one blue colony
which should lack lambda DNA, and three white colonies which
should contain phage sequences. The plasmids are then isolated
from the cultures by a unique mini-prep procedure that yields
high quality plasmid DNA without the use of toxic organic solvents.
Lab Session 4
Students digest the plasmids with EcoR1, BamH1, and a
combination of the two enzymes and determine the size of the
DNA fragments by electrophoresis. The results of this analysis
provide a first step in the identification of the lambda fragments
that were cloned.
Lab Session 5 - 7
The DNA fragments in the gels are transferred to nylon membranes
by the method of Southern using the transfer devices provided with
the program. The fragments containing the lambda sequences
are then detected by hybridization using a biotin-labeled probe
made from phage lambda DNA. The results of this analysis and
the restriction mapping studies performed during Lab Session 4
enable the student to identify the segment of the lambda genome
in each plasmid.
In 2012 we changed the Plasmid
Isolation procedure for this experiment.
s-ODERN"IOLOGY)NCss
25
Chemical Package Contents
Electrophoresis
Agarose
Gel Stain (Methylene Blue)
In Gel Stain
Electrophoresis Buffer
DNA Sample Buffer (2)
Dye Mixture
DNA Manipulation
Phage Lambda DNA
Plasmid pUC18 DNA
EcoR1 Endonuclease (2)
Bam H1 Endonuclease
Endonuclease Buffer (2)
DNA Ligase
Ligase Buffer
Bacterial Transformation and
Growth
E.coli
Calcium Chloride
Nutrient Broth (2)
Nutrient Broth + Ampicillin
Nutrient Agar + Ampicillin
X Gal
X Gal Solvent
E.coli-pUC18
Ampicillin
E.coli pUC18 – Lambda
Sterile Petri Dishes
Sterile Transfer Pipets
Sterile Microtubes
Blue
Colonies
In Standard Program 5, a library of
recombinant plasmids is prepared
by ligating DNA fragments from
phage lambda to linear pUC18. The
plasmids are introduced into E. coli
by transformation and white colonies
containing plasmids with lambda
DNA are recovered. Plasmids are
isolated from the E. coli and analyzed
by electrophoresis, and by Southern
blot hybridization procedures in order
to identify the segment of the lambda
genome contained in each. In the gel
shown, a plasmid from a blue colony
(lane 1), and plasmids from two
different white colonies (lanes 2 and
3), were digested with EcoR1 prior
to this electrophoretic analysis. DNA
standards are shown in lane 4.
1
2
3
4
DNA
White
Colonies
Sterile Culture Tubes
Inoculating Loops
Plasmid Isolation
(found in green box)
Solution I - 1 bottle
Solution II - 1 bottle
Solution III - 1 bottle
Equilibration Buffer - 1 bottle
Buffer HB - 1 bottle
Wash Buffer - 1 bottle
Elution Buffer - 1 bottle
HiBind DNA Column - 24
Collection Tubes - 24
RNase - 1 Tube
Southern Blot Analysis
Biotinylated Lambda DNA
4 Southern Transfer Devices
Nylon Membrane
Blotting Paper
Hybridization Bags
Hybridization Buffer
Tris Buffer
NP-40
Chloronapthol
Gelatin
Avidin – Peroxidase
Hydrogen Peroxide
Foam Rack
Paper Clips
Price List - Standard Laboratory Program 5
CAT. NO.
DESCRIPTION
S5
The Chemical Package for 16 students
working in pairs plus one student manual
and one instructor manual (The student manual
may be reproduced for educational purposes.)
S5-BK
8 Student Manuals
S5-SM
1 Student Manual and 1 Instructor Manual
PRICE
$297.46
$63.14
$9.61
For fast dependable service:
Call toll free: 800-733-6544
FAX: 765-523-3397
Visit: www.modernbio.com
&BTU4PVUIt-BGBZFUUF*/
s&!8s7EBSITEWWWMODERNBIOCOMs
26
Tissue to Gene
(Standard Laboratory Program 6)
A major goal of human genetics is to develop a complete map of the human genome. A high-resolution map of the
genome is considered to be essential for diagnosis of genetic diseases and development of cures of these diseases
by gene therapy. Studies on a specific DNA locus in a mammal usually begin with the isolation of genomic DNA
from a tissue. The DNA is quantified, subjected to restriction nuclease digestion, and the fragments separated by gel
electrophoresis. The DNA fragments in the gel are then transferred to a special membrane by the method of Southern
and the specific DNA fragment under investigation is detected by hybridization analysis. Your students will perform
each of these steps with this program series of five 3-hour laboratory sessions. The starting material for the series
is calf thymus tissue. Students isolate DNA from purified thymus nuclei and then quantify it with a novel solid phase
DNA assay. The DNA is digested with EcoR1, electrophoresed, and the separated fragments transferred to a nylon
membrane. The DNA fragment containing the G+C-rich cow satellite is then detected by hybridization using a biotinlabeled probe made from a plasmid that contains the cow satellite sequence. Students also determine if the satellite
sequence is found in sheep DNA. Microscopes, ethyl alcohol, an incubator that will maintain a temperature of 6065°C, and a centrifuge that can be operated at a force of at least 3000 x g are needed for the program.
DNA
Lab Session 1
Students isolate nuclei from thymus tissue and isolate thymus DNA
from the nuclei by a unique procedure that yields high quality
cellular DNA without the use of toxic organic solvents.
Lab Session 2
The purified DNA is quantified by a solid phase assay, digested with
the restriction endonuclease EcoR1, and then electrophoresed on
agarose gels.
We now sell extra nylon membrane
and blotting paper (folder).
This program takes 2 folders
Chemical Package Contents
Electrophoresis (ep-3/4)
Agarose
Gel Stain (Methylene blue)
In Gel Stain
Electrophoresis Buffer
Cow DNA (EcoR1 cut)
Sheep DNA (BamH1 cut)
DNA Sample Buffer
DNA Preparation
Calf Thymus Tissue
Cheese Cloth
Nuclear Buffer (2)
Sodium Dodecyl Sulfate
Ammonium Acetate (2)
Tris-EDTA Buffer
DNA Standard
Transfer pipets
Pasteur Pipets
EcoR1 (2)
Restriction Nuclease Buffer
Southern Blot Analysis
Foam Rack
Hybridization Buffer
NP40
Hydrogen Peroxide
Gelatin
Chloronapthol
Hybridization Bags
Nylon Membranes
Blotting Paper
Avidin-Peroxidase
Four Transfer Devices
Biotinylated Satellite DNA
Paper Clips
Lab Sessions 3-5
The DNA fragments in the gels are transferred to nylon membranes
using the transfer devices provided with the program. The fragment
containing the satellite sequence is then detected by hybridization
using a biotin-labeled probe prepared from a plasmid containing
the satellite sequence.
Price List - Standard Laboratory Program 6
CAT. NO.
DESCRIPTION
S6
The Chemical Package for 16 students
working in pairs plus one student manual
and one instructor manual (The student manual
may be reproduced for educational purposes.)
S6-BK 8 Student Manuals
S6-SM 1 Student Manual and 1 Instructor Manual
PRICE
$231.76
$63.14
$9.61
For fast dependable service:
Call toll free: 800-733-6544
FAX: 765-523-3397
Visit: www.modernbio.com
&BTU4PVUIt-BGBZFUUF*/
s-ODERN"IOLOGY)NCss
27
Plant Molecular Biology
(Standard Laboratory Program 9)
The chloroplast genome of higher plants is relatively small and simple. This genome contains about 120 genes
which code for chloroplast proteins. The organelle's genome also contains ribosomal RNA genes which code for
the RNA components of the chloroplast ribosome. In this program, students identify the ribosomal RNA genes in the
chloroplast genome of spinach. Students first isolate chloroplasts from fresh spinach and then prepare DNA from
the isolated organelle. The DNA is digested with EcoR1, electrophoresed, and the separated fragments transferred to
a nylon membrane. The DNA fragments containing the ribosomal genes are then detected by hybridization using a
biotin-labeled probe made from a plasmid that contains the ribosomal gene sequences. Fresh spinach, microscopes,
ethyl alcohol, a water bath incubator that will maintain a temperature of 60-65°C, and a centrifuge that can be
operated at a force of at least 3000 x g are needed for the program. A microcentrifuge is also desirable, but not
absolutely necessary.
Lab Session 1
Students isolate chloroplast from spinach leaves and isolate DNA
from the chloroplast by a unique procedure that yields high quality
DNA without the use of toxic organic solvents.
Lab Session 2
The purified DNA is digested with the restriction endonuclease
EcoR1, and then electrophoresed on agarose gels.
Electrophoresis (ep-3/4)
Agarose
Gel Stain (Methylene blue)
In Gel Stain
Electrophoresis Buffer
Lambda DNA (Hind III cut)
R-DNA Plasmid (EcoR1 cut)
DNA Sample Buffer
DNA Preparation
Large Transfer Pipets
Cheese Cloth
Buffer (Tris-Saline)
Sodium Dodecyl Sulfate
Ammonium Acetate
EcoR1 (2)
Restriction Nuclease Buffer
Southern Blot Analysis
Foam Rack
Hybridization Buffer
Tris Buffer
NP40
Hydrogen Peroxide
Biotinylated R-DNA Plasmid
Chloronapthol
Hybridization Bags
Blotting Paper
Nylon Membranes
Avidin-Peroxidase
Four Transfer Devices
Gelatin
Paper Clips
Agarose Gel Southern Blot
Extra nylon membrane
and blotting paper are
available on page 62.
Price List - Standard Laboratory Program 9
CAT. NO.
DESCRIPTION
S9
The Chemical Package for 16 students
working in pairs plus one student manual
and one instructor manual (The student manual
may be reproduced for educational purposes.)
S9-BK 8 Student Manuals
S9-SM 1 Student Manual and 1 Instructor Manual
Detection of Ribosomal RNA genes in Chloroplast DNA.
Left: Marker DNA and chloroplast DNA samples were electrophoresed on an
agarose gel and the gel was stained. Right: The separated DNA fragments on
the gel were transferred to a nylon membrane and the DNA containing the
chloroplast ribosomal RNA genes was identified by hybridization analysis.
A typical laboratory schedule for this program is given
where each lab session is approximately 3 hours.
s&!8s7EBSITEWWWMODERNBIOCOMs
PRICE
$231.19
$63.14
$9.61
DNA
Lab Sessions 3-5
The DNA fragments in the gels are transferred to nylon
membranes using the transfer devices provided with the program.
The fragments containing the genes for ribosomal RNA are then
detected by hybridization using a biotin-labeled probe prepared
from a plasmid containing the ribosomal gene sequences. Typical
results are shown below.
Chemical Package Contents
28
Cloning a DNA Segment from Sheep
(Standard Laboratory Program 11)
The sheep genome contains a highly repeated satellite sequence. In this program, students use recombinant DNA
techniques to clone this satellite DNA and then characterize the sequence by restriction enzyme mapping anaylsis.
The program provides essentially all of the instructions, chemicals, sterile media, and expendable accessories that
are needed to carry out DNA electrophoresis, restriction nuclease digestion, DNA ligations, bacterial transformations,
bacterial selection for ampicillin resistance and ß - glactosidase production and plasmid isolation. A microcentrifuge
is required for the program.
In 2012 we changed the Plasmid
Isolation procedure for this experiment.
DNA
Lab Session 1
Students digest sheep DNA with EcoR1 and electrophorese the
DNA fragments on an agarose gel in order to seperate the satellite
DNA from the remaining sequences in the sheep genome.
Lab Sessions 2-3
The satellite DNA band is eluted from the gel and ligated to EcoR1digested pUC18 in order to create a recombinant plasmid that
contains the satellite sequence. The recombinant plasmid is then
introduced into competent E.coli cells by transformation and
colonies containing foreign DNA are identified by a blue-white
color selection assay.
Lab Sessions 4-5
Students prepare overnight cultures from white colonies which
should contain the sheep satellite sequence. Plasmids are then
isolated from the cultures by a mini-prep procedure that yields
high quality plasmid DNA without the use of toxic organic solvents.
Students digest the plasmids with EcoR1 and determine the size
of the DNA fragments by electrophoresis. The results of this gel
analysis enable the student to identify the satellite sequence in the
recombinant plasmid.
For fast dependable service:
Call toll free: 800-733-6544
FAX: 765-523-3397
Visit: www.modernbio.com
&BTU4PVUIt-BGBZFUUF*/
Chemical Package Contents
Electrophoresis (ep-3/4)
Agarose
Gel Stain (Methylene Blue)
In Gel Stain
Electrophoresis Buffer
DNA Sample Buffer (2)
DNA Standards
DNA Manipulation
Sheep DNA
Plasmid pUC18 DNA (EcoR1 cut)
EcoR1 Endonulcease (2)
Endonuclease Buffer (2)
DNA Ligase
Ligase Buffer
DNA Elutor Components
1.5 ml tubes
0.5 ml tubes
Transfer Pipets
Glass Wool
Push Pins
Bacterial Transformation and
Growth
E.coli
Calcium Chloride
Nutrient Broth (2)
Nutrient Broth + Ampicillin
Nutrient Agar + Ampicillin
X Gal and Solvent
Ampicillin
Sterile Petri Dishes
Sterile Transfer Pipets
Sterile Microtubes
Sterile Culture Tubes
Inoculating Loops
Plasmid Isolation
(found in green box)
Solution I - 1 bottle
Solution II - 1 bottle
Solution III - 1 bottle
Equilibration Buffer - 1 bottle
Buffer HB - 1 bottle
Wash Buffer - 1 bottle
Elution Buffer - 1 bottle
HiBind DNA Column - 24
Collection Tubes - 24
RNase
Price List - Standard Laboratory Program 11
CAT. NO.
DESCRIPTION
S11
The Chemical Package for 16 student
working in pairs plus one student manual
and one instructor manual (The student
manual may be reproduced for educational
purposes.)
S11-BK 8 Student Manuals
S11-SM 1 Student Manual and 1 Instructor Manual
PRICE
$224.12
$63.14
$9.61
Find Equipment, Accessories, and Kits for
Teaching and Research starting on page 48.
s-ODERN"IOLOGY)NCss
29
Mini Programs for DNA Analysis
Watch a video of this experiment in
action at MODERNBIO.COM
IND-9. Producing a Strain of E.coli that Glows in the Dark
The emission of light by living organisms is a fascinating process.
The genetic system required for luminescence in the bacterium
Photobacterium (Vibiro) fischeri is the lux operon. This operon
contains a gene for luciferase (the enzyme that catalyzes the light-emitting
reaction) and genes for enzymes which produce the luciferins (which
are the substrates for the light-emitting reaction.). In this exercise,
students create a luminescent population of bacteria by introducing
into E.coli a plasmid that contains this lux operon. The success of the
transformation is readily apparent since the E.coli colonies that take
up this plasmid glow in the dark as shown on the right. The simple
procedure can be carried out during a single 1-1.5 hour laboratory
CONTROL PLASMID
PLASMID LUX
Light
The photograph in the
bottom panel was taken
in total darkness using
only the light emitted
from the bacteria that
were transformed with
plasmid lux.
Dark
PROCEDURE AND RESULTS
1. Add E.coli to calcium chloride solution.
2. After 15 minutes, add control plasmid and plasmid lux to two
tubes of cells and incubate both on ice for10 minutes.
3. Incubate the tubes at 37°C, add nutrient broth and then spread the
bacterial suspensions onto agar plates.
4. View the plates in the light and in the dark after two days at room
temperature.
CAT. NO.
DESCRIPTION
PRICE
IND-9
Producing a Strain of E.coli that Glows in the Dark
$84.21
Includes instructions, E.coli, plasmid lux, control plasmid, calcium
chloride, inoculating loops, transfer pipets, tubes, petri dishes,
ampicillin,nutrient agar-ampicillin, and nutrient broth. All
materials are provided sterile and ready for use.
DNA from the normal rabbit ß-globin
gene (odd numbered lanes) and DNA from
a deletion mutation of the gene, (even
numbered lanes) were digested with
EcoR1 (lanes 1, 2, 5, 6) or Hae III (lanes 3, 4,
7, 8) prior to this electrophoretic analysis.
The mutant lacks DNA fragments which
are found in the normal gene.
This experiment can be
used in a 1 hour class.
DNA
Electrophoresis
equipment not
required.
IND-6. Analysis of a Mutant Hemoglobin Gene
A mutation is a change in the nucleotide sequence of DNA
which leads to an inherited change in an organism. Restriction
endonucleases provide valuable tools for characterizing mutations
at the DNA level. This principle is illustrated in the exercise
where students digest a normal and a mutant gene with EcoR1
and Hae III and then analyze the DNA fragments from each by
electrophoresis as shown in the figure on the right. The gene is
from rabbit and codes for the ß-globin chains of hemoglobin.
1 2 3 4 5 6 7 8
session. Sufficient sterile materials are provided for sixteen platings and
the experiment is designed for 8 groups of students.
Additional experiments can be performed following completion of this
experiment including:
1. Analysis of the effects of growth temperature and time on the
glowing process. This analysis can be carried out by replating
the transformed cells onto fresh nutrient agar-ampicillin plates,
Nutrient agar can be purchased separately (see page 62).
2. Isolation of plasmid lux from transformed cells using the plasmid DNA
isolation kit (cat. # 3-18) that is described on page 62.
The results of the analysis enable the student to identify the
type of mutation and the position of the mutation in the globin
gene. The exercise was designed for eight groups of students
and is provided in one of two forms. In IND-6A, the DNAs have
been predigested with restriction enzymes and are ready for
electrophoresis. In IND-6B, students digest the DNA with restriction
enzymes prior to electrophoretic analysis. Electrophoresis
Package 3/4 (Page 21) is needed, but not included.
CAT. NO.
DESCRIPTION
PRICE
IND-6A Predigested DNA for agarose gels
$66.27
Includes instructions and plasmids containing normal and mutant
rabbit ß-globin genes predigested with EcoR1 and Hae III.
CAT. NO.
DESCRIPTION
PRICE
IND-6B Nondigested DNA for agarose gels
76.54
Includes instructions and plasmids containing normal and mutant
rabbit ß-globin genes, EcoR1, Hae III., restriction nuclease buffers
and electrophoresis sample buffer.
s&!8s7EBSITEWWWMODERNBIOCOMs
D
N
A
30
Mini Programs for DNA Analysis
IND7. AMPLIFICATION OF A HEMOGLOBIN GENE BY THE POLYMERASE CHAIN REACTION PCR
DNA
The polymerase chain reaction (PCR) is one of the most powerful
techniques used in molecular biology. With this method, a few
ng of DNA can be amplified millions of times in a test tube in a
few hours. The PCR has been used extensively in studies of gene
structure and function. The method is also becoming increasingly
important in DNA typing procedures such as DNA fingerprinting
and in the identification and characterization of mutations that
cause human diseases. This exercise is designed to illustrate
the PCR in the teaching laboratory. In the experiment, students
use PCR to amplify a rabbit ß-globin gene. The template for this
reaction is a plasmid that contains this globin sequence. Following
PCR amplification, the product of the reaction is analyzed
on an agarose gel as shown on the right. The experiment was
designed so that the PCR can be done manually provided that
three standard water baths (45°C, 74°C, and 94°C) or three hot
plates are available. The amplified globin DNA band can clearly
be seen following staining of the
agarose gels with methylene blue.
The amplification reaction requires
approximately 1.5 hours. Sufficient
materials are provided so that 8
groups of students can perform the
experiment. The following materials
are needed but not included: Three
water baths (or an authorized DNA
Thermal Cycler), electrophoresis
equipment and Electrophoresis Package
3/4 (See page 21).
1
2 3
4 5 6
7 8
Rabbit ß-globin genes
were amplified by PCR and
electrophoresed on lanes 4 and
7 of this agarose gel. Lanes 2
and 5 show DNA size markers.
CAT. NO.
DESCRIPTION
PRICE
IND-7 Amplification of a Hemoglobin Gene by the
$124.91
Polymerase Chain Reaction (PCR)
Includes instructions , PCR buffer, Taq DNA polymerase, DNA
template, primers, nucleotides, Taq dilution buffer, DNA size markers,
electrophoresis sample buffer, mineral oil, and PCR tubes.
IND10. PCR AMPLIFICATION OF A GENE FOR RIBOSOMAL RNA FROM DIFFERENT ORGANISMS
A ribosome is composed of a large subunit and a small subunit
and each subunit contains specific RNAs that are associated with
ribosomal proteins. Ribosomes play the same essential role in
protein synthesis in all contemporary organisms and the genes
for ribosomal RNAs in each subunit are highly conserved. The
gene for the small-subunit ribosomal RNA (16 S-like) has been
especially useful in evolutionary studies of distant phylogenetic
relationships because it has remained quite stable during
evolution of all organisms. The 16 S-like ribosomal genes can
be amplified by the PCR from total DNA isolated from essentially
any organism by using a single set of primers that recognize
conserved regions of the gene. In this exercise, students use the
PCR to amplify the gene for the 16 S-like ribosomal RNA from cow,
pig, chicken and E.coli. The templates that are provided for this
experiment are genomic DNAs from these organisms. In addition
to these templates, students may amplify 16 S-like ribosomal
genes using genomic DNAs that they have isolated from bacteria,
plants or animals. These genomic DNAs can be isolated by using
the genomic DNA isolation kit (Cat.# 3-19) described on page
62 of this catalog. The amplified 16 S-like ribosomal RNA genes can
clearly be seen as discrete bands following staining of the agarose gels
*The Polymerase Chain Reaction is covered by patents owned by Hoffman-LaRoche, Inc.
The exercise described above is to be used for teaching purposes only.
with methylene blue as shown on the right. The experiment was designed
so that the PCR can be done manually provided that three standard water
baths (45°C, 74°C, and 94°C) or three hot plates are available. The
amplification reaction requires
approximately 2 hours. Sufficient
materials are provided so that 8
groups of students can perform
the experiment with each group
amplifying the ribosomal RNA gene
from one template. The following
materials are needed but not
included: Three water baths (or an
authorized DNA Thermal Cycler), I6 S - like ribosomal genes were
by PCR using genomic
electrophoresis equipment and amplified
DNA from seven different
Electrophoresis Package 3/4. (See organisms. DNA size markers are
shown in the first lane.
page 21)
CAT. NO.
DESCRIPTION
PRICE
IND-10 PCR Amplification of a Gene for Ribosomal
$162.82
RNA from Different Organisms
Includes instructions, PCR buffer, Taq DNA polymerase, DNA
templates (cow DNA, pig DNA, chicken DNA, E.coli DNA), primers,
nucleotides, Taq dilution buffer, DNA size markers, electrophoresis
sample buffer, mineral oil, and PCR tubes.
s-ODERN"IOLOGY)NCss
31
IND21. IDENTIFYING GENOMIC AND PLASMID
DNA SEQUENCES IN E.COLI BY COLONY PCR
Purified DNA is most often used as a template in the PCR reaction.
However, it is possible to amplify specific DNA sequences without
DNA purification by starting with a single living E.coli colony. This
technique is known as colony PCR and provides a powerful and
reliable method for the rapid amplification and isolation of any
gene in the E.coli genome or any gene on a plasmid that is carried
by E.coli. In this exercise, students carry out colony PCR starting
with a culture of E.coli that carries an ampicillin-resistance gene
on plasmid pUC18. They first streak the cells over an nutrient agar
plate in order to produce single E.coli colonies. These colonies
are then used directly as templates in a PCR reaction in order
to amplify a segment of the 16 S -ribosomal RNA genes which is
located on the E.coli chromosome and a segment of the ampicillinresistance gene on the plasmid. Sufficient materials are provided
so that 8 groups of students can perform the experiment with
each group amplifying the ribosomal RNA gene, the ampicillinresistance gene or both genes. Typical results of the experiment
are shown on the right. A DNA Thermal Cycler, electrophoresis
equipment and Electrophoresis Package 3/4 (See page 23) or
equivalent are needed but not included.
16S-ribosomal RNA genes which are located on the E. coli
chromosome and the ampicillin-resistance gene on a plasmid
were amplified by colony PCR using a single E. coli colony as
template. From left to right, samples are DNA markers, the
16 S -ribosomal RNA gene,the ampicillin-resistance gene and
both genes.
CAT. NO.
DESCRIPTION
PRICE
IND-21 Identifying Genomic and Plasmid DNA
Sequences in E.coli by Colony PCR
$168.39
Includes instructions, E. coli-pUC 18, nutrient agar-ampicillin,
ampicillin, petri dishes, inoculating loops, ampicillin-resistance
gene primers, rDNA primers, sterile pipet tips, PCR buffer,Taq
polymerase, Taq dilution buffer, nucleotides, DNA size markers,
electrophoresis sample buffer, mineral oil, and PCR tubes.
DNA
IND12. CHARACTERIZATION OF THE SATELLITE DNA FROM THE MEAL WORM
Satellite DNAs are highly repeated sequences of unknown function.
The satellite DNA from the meal worm beetle is remarkable since
it represents over 50% of this insects genome. In this exercise,
students first isolate DNA from beetle larvae by a simple and safe
procedure. They then digest the DNA with EcoR1 and examine the
satellite DNA following electrophoresis as shown on the gel at the
right. Sufficient materials are provided so that the experiment can
be carried out twice by eight groups of students. The exercise can
be completed in two, 2 to 3 hour laboratory sessions and requires
ethyl alcohol, a microcentrifuge and Electrophoresis Package 3/4
(see page 21).
In this exercise, DNA isolated from beetle larvae was digested
for 0, 5, and 20 minutes (lanes 1-3, 5-7) with EcoR1 prior to this
electrophoretic analysis. Note the progressive breakdown of the
DNA satellite to the satellite monomer which is 142 base-pairs
in length. DNA size markers are shown in lanes four and eight.
1 2 3 4 5 6 7 8
CAT. NO.
DESCRIPTION
PRICE
IND-12 Characterization of the Satellite DNA from
$75.52
the Meal Worm
Includes instructions, meal worm beetle larvae, EcoR1(2), EcoR1
buffer, extraction buffer, SDS, ammonium acetate, transfer pipets,
sample buffer, and DNA Standards.
s&!8s7EBSITEWWWMODERNBIOCOMs
32
Electrophoresis
equipment not
required.
IND13. TRANSFORMATION
AND ANALYSIS OF YEAST
DNA
The budding yeast Saccharomyces cerevisiae has served as one
of the most important model organism for the study of eukaryotic
molecular biology. The entire yeast genome has been sequenced
and the function of many yeast genes is now known. This rapid
progress has relied heavily on the ability to introduce replicating
plasmids into yeast. A typical replicating yeast plasmid contains
an origin of replication known as an autonomously replicating
sequence or ARS and a selectable marker gene that enables the
yeast to grow in the absence of a nutrient such as uracil. In this
exercise, students introduce such a plasmid into yeast and then
select for transformants by plating the cells on a nutrient-agar
medium lacking uracil. They then study the stability of the plasmid
in yeast cells by growing the cells in the presence of uracil. The
program lasts two, 2 to 3 laboratory sessions and requires
basic training in microbiology and was designed for 8 groups of
students. The following materials and equipment are needed but
not included: microcentrifuge, table-top centrifuge, autoclave to
prepare media, bacterial spreading rod, Bunsen burners, sterile
water and microliter dispensers.
CAT. NO.
DESCRIPTION
PRICE
IND-13 Transformation and Analysis of Yeast
$190.68
Includes instructions, plasmid DNA, carrier DNA, lithium acetate,
polythylene glycol, 10 x TE buffer, dehydrated nutrient agar
minus uracil, dehydrated nutrient broth minus uracil, uracil,
sterile microtubes, sterile culture tubes inoculating loops,
and uracil plus and uracil minus yeast cultures.
Electrophoresis
equipment not
required.
IND15. BACTERIAL
PHENOTYPE
Genes introduced into E.coli by plasmid-mediated transformation
can confer variation in the phenotype of the bacteria. For example,
E.coli containing the ampicillin-resistance gene grows in the
presence of this antibiotic while the product of the ß-galactosidase
gene enables the bacteria to convert the ß-galactosidase substrate
X-gal to a blue product. Similarly, E.coli containing the Lux
operon produce colonies that glow in the dark. In this exercise,
four different strains of E.coli are streaked onto agar plates
containing media that selects for the different strains and that
reveals variation in colony phenotype. Students record these
variations and the results enable them to deduce the genotype
of the strains as shown in the Table below. Sufficient sterile
materials are provided to prepare 32 plates and the experiment
was designed for 8 groups of students. The exercise requires two
1 hour laboratory sessions.
CAT. NO.
DESCRIPTION
PRICE
IND-15 Includes instructions, four strains of E.coli
$82.69
(A-D: teacher keyed), inoculating loops, nutrient agar, petri
dishes, ampicillin, X-gal, X-gal solvent.
Colony Phenotype
E.coli
Strain
A
B
C
D
(SPXUIPO
Ampicillin
No
Yes
Yes
Yes
$PMPOZ$PMPS
on X-Gal Plates
White
Blue
White
(MPXTJO
the dark
No
No
Yes
Genotype
Gene(s) Responsible
GPS$PMPOZ1IFOPUZQF
Ampicillin Resistance
Ampicillin Resistance+Galactosidase
Ampicillin Resistance+Lux
IND28. SYNTHETIC BIOLOGY: USING BACTERIAL COMPUTERS TO SOLVE THE PANCAKE PROBLEM
Synthetic biology is an exciting new field that uses engineering
principles and mathematical modeling to design and construct
biological devices. Synthetic biology projects include the
construction of bacterial computers that can solve mathematical
problems. Microbial machines, in the form of genetically
engineered E. coli cells, have solved a variety of mathematical
problems, which have had important applications in biology,
medicine and technology. This investigation introduces students to
synthetic biology principles as they use genetic recombination in
bacterial computers to solve the Burnt Pancake Problem. In the
project, the biological equivalent of a burnt pancake is a functional
module of DNA. Similar to stacks of pancakes burnt on one side,
DNA modules have directionality, require a specific order and
can be flipped by genetic recombination. The bacterial computer
is investigated using a modular system in which pancake stacks
are assembled from flippable DNA segments. Flipping of the DNA
segments "pancakes" is performed by a DNA recombination system
and the flipping is monitored by antibiotic sensitivity of the bacteria
and colony color. The investigation interlaces biology laboratory
work with mathematical modeling, thereby allowing students
to explore the rich interface between biology and mathematics.
Sufficient sterile media, antibiotics, cells, plasmids Electrophoresis
and other materials are provided so that 16
equipment
not required.
students working in pairs can perform the analysis.
CAT. NO.
DESCRIPTION
IND-28 Synthetic Biology: Using Bacterial Computers
to Solve the Pancake Problem
PRICE
$180.00
Panel A shows the starting configuration of the burnt pancake bacterial computer. The
double helix figures indicate the three recombination sites for a specific recombinase
called Hin. The two DNA segments between these sites define the pancakes. Depending
on the orientation of these segments, the gene for the red fluorescent protein (RFP) and
tetracycline resistance gene (Tet) are either active or inactive and these activities are
monitored by the student to deduce these orientations. Panel B shows the differential
expression of the RFP in different bacterial clones as a result flipping induced by the
recombinase. Panel C shows the activities carried out during the 5 laboratory sessions
that comprise this exploration.
s-ODERN"IOLOGY)NCss
33
Contemporary Cell Biology
Standard Laboratory Program 7
Today’s cell biology is a fascinating blend of molecular biology, biochemistry, and a variety of traditional disciplines. These
fields are integrated into this program in order to introduce students to selected topics in eukaryotic cell biology. In the
program, students localize enzymes in plant and animal cells, perform cell fractionation procedures, and study the properties
of a specific membrane receptor. Each exercise can be completed in one 3-hour or two 90-minute laboratory sessions.
Microscopes, slides, microliter dispensers, small test tubes, tabletop centrifuge, and ethyl alcohol are needed but not provided.
701. Enzyme Cytochemistry
Electrophoresis
equipment not
required.
EXPERIMENTS
The concept that different enzymes are found in different tissues, cell
types, and cell organelles is illustrated in this multipart exercise where
students use contemporary techniques to localize specific enzymes in
cells and tissues. Students are introduced to enzyme cytochemistry in an
experiment on the germinating corn seed where they show that peroxidase
is produced by the aleurone, a cell layer that surrounds the endosperm.
They then characterize the subcellular distribution of peroxidase in
giant onion epithelial cells and show that the enzyme resides in the cell
wall. In the final experiment of the series, the subcellular distribution
of lactate dehydrogenase in chicken erythrocytes is examined where
students learn that this enzyme is confined to erythrocyte cytoplasm.
Concanavalin A-Induced Hemagglutination
- Mannose
+ Mannose
Concanavalin A Binding
- Mannose
+ Mannose
702. Analysis of a Cell-Surface Receptor
Chemical Package Contents
EXP-701
Chicken Blood Smears
Plastic Trays with Lids
Tris Buffer
Methylene Blue
Hydrogen Peroxide
Chloronapthol
Tetrazolium
Iodine Solution
Transfer Pipets
Corn Seed
Eosin Transfer Pipets
LDH-Substrate
NP-40
EXP-702
Hydrogen Peroxide
Tris Buffer
Con A
Con-A-Peroxidase
Mannose
Con-A-Buffer
Galactose
Transfer Pipets
EXP-703
DNAse I
Chloronapthol
Methylene Blue
Eosin
Onion Bulbs
Calf Thymus
Nuclear Buffer
Hydrogen
Peroxide
Tetrazolium
Tris Buffer
SDS
5M NaCl
The results of EXP 702 show that mannose selectively inhibits the concanavalin A-induced
hemagglutination reaction and the binding of concanavalin A to receptors on the surface of
cheek epithelial cells.
703. The Cell Nucleus
The exercise is introduced with a discussion of observations that were
made in the early nineteenth century. Namely, that biological stains often
show specific affinity for particular subcellular components. With this
information, students use differential staining procedures to identify
nuclear and cytoplasmic components in cells from thymus and onion
root tip. In the second part of the exercise, cell nuclei are isolated
from thymus tissue and their purity assessed by staining techniques.
The DNA is isolated from the nuclei by a procedure designed to
provide information on the packaging of this macromolecule in
the chromosome. The exercise gives students experience with both
classical and contemporary techniques, and makes them aware that
cell and molecular biology have long histories that can be traced to
traditional disciplines.
STANDARD LABORATORY PROGRAM 7
CAT. NO.
DESCRIPTION PRICE
S7
The Chemical Package for 16 students plus one
204.50
student manual and one instructor manual.
S7-BK 8 Student Manuals
$63.14
S7-SM Sample Student Manual (47 pages) plus one
$9.61
instructor manual.
INDIVIDUAL EXPERIMENTS
EXP-701 Enzyme Cytochemistry
EXP-702 Analysis of a Cell-Surface Receptor
EXP-703 The Cell Nucleus
s&!8s7EBSITEWWWMODERNBIOCOMs
$71.77
$84.12
$71.77
DNA
Chemical signaling between cells in multicellular organisms is
frequently mediated by cell-surface receptors. The receptors for
neurotransmitters, protein hormones, growth factors, and plant lectins
are a few of the many known examples of these important membrane
components. In this exercise, students examine the cell location and
properties of the receptor for the lectin concanavalin A. In the first
experiment of the series, students use a concanavalin A-peroxidase
complex in a microscopic assay to show that the specific receptor is
found on the surface of their own cheek epithelial cells. In the second
part of the exercise, students study the characteristic hemagglutination
reaction that is elicited by concanavalin A and learn that the reaction
is due to multiple sites for receptor binding on the concanavalin A
molecule. It is known that concanavalin A binds selectively to mannose
residues on the cell-surface glycoprotein receptor. This specificity is
confirmed in the final experiment of the series where students show that
mannose but not other sugars inhibit concanavalin A binding and the
hemagglutination reaction. This innovative exercise provides a detailed
picture of a cell-surface receptor and presents strategies that are used
to study this important class of regulatory molecules. Typical results of
selected portions of the experiment are shown on the right.
34
Mini Programs in Cell Biology
IND2. TISSUE PRINTING
Electrophoresis
equipment not
required.
Locating specific proteins and nucleic acid molecules in tissue sections
is an important goal in cell biology. An effective and simple technique for
this purpose is tissue printing which permits the localization of specific
macromolecules in animal and plant tissues. Here students perform this
technique to examine the tissue distribution of the enzyme peroxidase
in plants. First, students section carrots, celery, and other vegetables
with razor blades and transfer the proteins from the cut sections to
nitrocellulose membranes by application of gentle pressure. The
enzyme peroxidase is detected on the membranes using a blue color
producing peroxidase substrate as shown on the left. All proteins on
the membranes are then stained red with a general protein stain which
highlights the selective nature of the peroxidase distribution. Students
also determine the amount of peroxidase in plant extracts. This 2 hour
exercise provides an exciting lesson in plant histology and introduces
your student to an important technique used in contemporary cell
biology. The exercise was designed for eight groups of students and
includes: nitrocellulose membranes, transfer pipets, 200 x extraction
buffer, peroxidase standard, chloronapthol, 4M Tris buffer, hydrogen
peroxide, and 10 x protein blot stain. Assorted vegetables which can be
purchased at your local supermarket and petri dishes are needed, but
not provided.
Cross-sections of carrots (large circles), parsnips (small circles), and celery
(horseshoe-shaped) were prepared with a razor blade. The proteins at the
cut surface of the sections were transferred to nitrocellulose membranes by
application of gentle pressure for 10 seconds. The nitrocellulose was then
incubated with a color producing peroxidase substrate. Note that the peroxidase
is found primarily in the epidermis and in some vascular (phloem) elements.
CAT. NO.
DESCRIPTION
IND-2 Tissue Printing
PRICE
$73.84
CELL BIOLOGY
IND3. THE ELISA IMMUNOASSAY
The ELISA (enzyme-linked immunosorbant assay) is a powerful
immunological method for detecting specific proteins in complex
protein mixtures. The ELISA has become an important tool for the
cell and molecular biologist. It is increasingly being applied in clinical
medicine for detecting proteins associated with disease including
antibodies produced in response to infection by the HIV virus. The
method is also easy to perform and yields graphic results making it well
suited for the teaching laboratory. In this exercise, students carry out
the ELISA to study the specificity of antibody-antigen interactions. The
basic procedures and typical results of the exercise are shown on the
right. The 2-3 hour exercise was designed for sixteen students working
in pairs and includes: microtitration plates, transfer pipets, chicken
serum, cow serum, horse serum, rabbit serum, gelatin, tris buffer
saline, tris buffer saline + NP-40, goat anti-rabbit IgG-peroxidase, TMBperoxidase substrate, and hydrogen peroxide. Microliter dispensers
are needed but not provided, and a colorimeter is desirable but not
Electrophoresis
absolutely necessary.
equipment not
required.
CAT. NO.
DESCRIPTION
IND-3 The ELISA Immunoassay
PRICE
$73.84
Serum Concentration
1. Varying dilutions of serum proteins from chicken (D and H), cow (C and
G), horse (B and F), and rabbit (A and E) are adsorbed to the surface of wells
of a microtitration plate. The antigen used for the experiment is the IgG
molecule from rabbit serum.
2. The immobilized serum proteins are incubated with a peroxidase-linked
antibody to the rabbit antigen. During this incubation, the antibody binds
preferentially to the antigen in rabbit serum.
3. The bound antibody is detected by a color-producing reaction and the
amount of color development is proportional to the amount of antigen
present. Note maximal color development occurs with the rabbit serum
proteins.
s-ODERN"IOLOGY)NCss
35
IND22. CHARACTERIZATION OF PEROXIDASES IN PLANTS: STUDENT DESIGNED RESEARCH PROJECTS
This program was designed to actively engage students in exciting biological
research projects of their own design. The projects focus on peroxidases,
which form a large family of related enzymes that are ubiquitous in plants.
Plant peroxidase isoenzymes can be tissue specific, developmentally
regulated and display variable tissue and, high salt and disease resistance
defense reactions. This induction may be related to the abilities of
peroxidases to strengthen the plant cell wall and to kill microorganisms.
Students begin their projects with a hypothesis, which is a statement of an
ideal that they will test in the laboratory. They then test their hypothesis by
carrying out a series of experiments using the materials provided with the
program and vegetables, intact plants or roots and stems. They first use the
technique of tissue printing which enables them to localize the peroxidases
in tissue sections. They then quantify peroxidase activity by using a DOTblot assay and spectrophotometric procedures. Students also carry out an
electrophoresis analysis in order to characterize peroxidase isoenzymes in
plant extracts that they prepare. In the final section of the program, students
are given detailed instructions for organizing their data for presentation in a
scientific paper. They are then instructed to write a paper that conforms to
the style of a scientific publication using the detailed steps that are presented
in the laboratory manuals. This program for requires about three 2-3
hour laboratory sessions. Electrophoretic equipment, spectrophotometer,
and plants/vegetables are needed but not provided. The materials that are
provided for the program and a few of the nearly infinite number of titles of
student papers are given on the right.
Extra nitrocellulose packets
available. See page 62 for details.
Chemical Package Contents
Electrophoresis
Agarose
Electrophoresis Buffer
Sample Buffer
HRP-Basic
HRP-Mixture
Cytochrome C
Serum Albumin-Hemoglobin
Peroxidase Assays
Hydrogen Peroxide (3)
Chloronapthol (5)
200 X Extraction Buffer (3)
Color Development Buffer
Peroxidase Standard (3)
10 X Protein Blot Stain
Nitrocellulose (4)
Transfer pipetes
A FEW TITLES OF STUDENT PAPERS
s %FFECTSOFSALTSTRESSONTHELEVELSTISSUEDISTRIBUTIONAND
isoenzyme profiles of peroxidases in the turnip.
s $EVELOPMENTAL CHANGES IN THE CORN STEM AS VIEWED BY
PEROXIDASETYPESAMOUNTSANDTISSUELOCATION
s 6ARIATIONS IN PEROXIDASE LEVELS TISSUE DISTRIBUTION AND
isoenzymes in different regions of the carrot.
CAT NO.
DESCRIPTION
IND-22 Characterization of Peroxidases in Plants
PRICE
$139.21
IND29. ELECTROPHORETIC AND CHROMATOGRAPHIC ANALYSIS OF
PHOTOSYNTHETIC PIGMENTS FROM BLUE GREEN ALGAE
show that it contains the single major protein Phycocyanin by
electrophoresis as shown in the gel below. They also determine
the charge of this protein by comparing its electrophoretic
mobility to the mobilities of dyes with known charges. In part B,
they prepare an alcohol extract and analyze the smaller alcohol
soluble pigments by thin layer chromatography in order to identify
the chlorophylls and major carotenoid pigments. The results of
this two-part study give students practical hands-on experience
with isolation of components from cells as well as electrophoresis
and thin layer chromatography and introduces them to one of the
most important organisms on the earth.
Chemical Package Contents
What's Included
Complete Instructions
Blue –Green Algae
Dye Sample 2
Dye Sample 2
Glycerol Solution
Thin-Layer Sheet
Chromatographic Tubes
1.5 ml Tubes
1ml Transfer Pipits
s3MALL0IPITS
Bovine Hemoglobin
Accessories and Chemicals Needed
Electrophoresis Equipment
Automatic Pipetors (Optional)
Methanol
CAT NO.
DESCRIPTION
IND-29 Electrophoretic and Chromatographic Analysis of
Photosynthetic Pigments from Blue- Green Algae
s&!8s7EBSITEWWWMODERNBIOCOMs
PRICE
$65.28
CELL BIOLOGY
Cyanobactera, also known as blue-green algae, obtain their
energy by photosynthesis using sunlight as their energy source.
These organisms have been considered to be the oldest and
the most important bacteria on the earth. It is believed that
they were responsible for the initial oxygenation of the earth's
atmosphere through photosynthesis and it is also felt they were
the precursors to the chloroplasts that are found in true algae
and plants. There are two classes of photosynthetic pigments
in Cyanobactera. The first class contains water-soluble proteins
and the major protein is called Phycocyanin, which is blue.
The other classes of photosynthetic pigments that include the
carotinoids and chlorophylls
are small molecular weight
molecules and are insoluble
in water but soluble in organic
solvents such as alcohol. In
this laboratory exercise, 8
groups of students isolate
and characterize both groups
of pigments. In part A of this
exercise, students prepare
a
water-soluble
extract
from blue green algae and
36
Mini Programs in Cell Biology
IND17. A RAPID IMMUNOLOGICAL
METHOD TO STUDY EVOLUTION
CELL BIOLOGY
Each protein carries in its amino acid sequence information
pertaining to its evolutionary history and origin, and provides
clues to the evolutionary history of the organism in which it is
found. Indeed, proteins existing today are in effect living fossils.
This concept is illustrated in this exercise where eight groups of
students examine the abilities of antibodies against cow gamma
globulin to react with gamma globulins in the sera of cow, goat,
sheep, horse, and chicken. The results of the experiment will
enable the student to answer the questions on the right which are
posed in the Student Guide provided with the exercise. Answers
to these questions are given in the Instructor Guide. The method
used for the experiment involves dotting small quantities of serum
onto nitrocellulose and then incubating the nitrocellulose with an
enzyme-linked antibody against cow gamma globulin. Following
development of the nitrocellulose, purple dots appear with
intensities that are proportional to the reactivity of the antibody.
The entire exercise was designed for eight groups of students and
can be completed within a 2-hour laboratory period. Microliter
dispensers are needed but not provided and electrophoresis
equipment is not required.
Chemical Package Contents
8 sheets of nitrocellulose
blotting paper
dishes for nitrocellulose incubation
anti-Cow IgG (Peroxidase linked)
purified cow IgG
cow serum
horse serum
goat serum
sheep serum
chicken serum
protein blot stain
hydrogen peroxide
chloronapthol
gelatin
Tris buffer
Tris buffer saline (TBS)
TBS+NP-40
Electrophoresis
equipment not
required.
Study Questions and Data Analysis
1. Indicate the tree (A, B, C or D) shown below which is consistent
with the results of your experiment and explain the reasons for
your choice.
Sheep
A
Horse Goat
Goat
Cow
B
Sheep
Horse
Cow
Chicken
Chicken
Reptiles
Reptiles
C
D
Cow
Sheep Horse
Goat
Chicken
Sheep
Goat
Cow
Chicken
Horse
Reptiles
Reptiles
2. Does your answer given above agree with what is known about
the phylogeny of these vertebrates?
3. Construct a standard curve for the gamma globulin by plotting
the concentration of purified gamma globulin against relative spot
intensities. Using this curve, estimate the concentration of gamma
globulin in the cow serum sample.
CAT. NO.
IND-17
DESCRIPTION
A Rapid Immunological Method to Study Evolution
PRICE
$84.86
IND14. ENZYME KINETICS
This miniprogram was designed to give students a basic
calculate values of Vmax and Km. The series can be completed in
understanding of enzyme kinetics. In the first experiment in this two 2-3 hour laboratory sessions and requires a colorimeter or
Electrophoresis
series, students prepare an extract from wheat germ. They then spectrophotometer and a table-top centrifuge.
equipment not
determined the initial velocity (Vo) of the reaction catalyzed by
required.
purified acid phosphatase and by the acid phosphatase activity
CAT. NO.
DESCRIPTION
PRICE
present in the extract. From these data, they estimate the amount
IND-14 Enzyme Kinetics
$76.78
Includes instructions, wheat germ, acid phosphatase,
of the enzyme that is present in the wheat germ. In the second
extraction buffer, acid phosphatase substrate, substrate
experiment, the student examines the effects of substrate
dilution buffer, nitrophenol standards, small transfer pipets
and large transfer pipets.
concentration on the reaction velocity. Data are analyzed using
Michaelis-Menton and Lineweaver-Burk plots and students
s-ODERN"IOLOGY)NCss
37
Zoology Histology Developmental Anatomy
College students are frequently introduced to basic histology, zoology and developmental anatomy during their
freshman and sophomore years. Traditionally, this is accomplished by the study of prepared microscope slides. The
approach does not teach students how slides are made and students frequently become disinterested and bored
because they are viewing prepared biological materials. Modern Biology Inc. now offers three miniprograms that
were designed to circumvent these limitations. The starting point for the programs are precut tissue sections which
are ready for staining. Thus, expensive slide–making equipment is not required. During the first 2-3 hour laboratory
session, students process the sections through stains (hemotoloxin and eosin) and prepare permanent slides of
the tissues. A nontoxic and biodegradable histological clearing agent is used in the procedure which eliminates
hazards associated with the use of xylene or toluene-based clearing agents. During the second and third laboratory
sessions, students follow the well-illustrated laboratory guides to receive a fine introduction to histology, zoology or
developmental anatomy. Each of these programs is an independent teaching unit and is designed for a class of 16
students working in pairs. Sufficient materials are provided so that 8 groups can make 2 slides each. Microscopes and
forceps are needed but not provided. Although the programs were designed to integrate three 2-3 hour laboratory
sessions in classical biology into freshman or sophomore–college level biology classes, they can also can be used to
supplement more advanced courses in histology, developmental biology and zoology.
IND19. AN INTRODUCTION TO
VERTEBRATE HISTOLOGY
Molecular and cell biology can be understood and
appreciated only in the light of their relations to the
whole
organism.
However, in recent
years it has become
fashionable to reduce
or to eliminate
altogether laboratory
time allotted for
organismic biology.
In this program,
students
prepare
and then study
cross sections of
planaria, earthworm, lamprey (larvae), and tadpole.
The laboratory guides emphasize hierarchies of animal
complexity and the similarities and differences between
the invertebrates and vertebrates. The picture above
shows a student-prepared slide of the earth worm.
Students prepare and then study sections of
trachea and esophagus in order to identify
and characterize the major tissue types (
repithelia, connective tissue and muscle).
They then study selected organs including
ovary, testis, intestine, spinal cord and skin.
The picture above shows a student-prepared
slide illustrating the trachea and esophagus.
Chemical Package Contents
Chemical Package Contents
Planarian Slides (4)
Earthworm Slides (4)
Lamprey (larvae) Slides (4)
Tadpole Slides(4)
Hematoxylin
Eosin Y
Isopropanol (2)
Esophagus/Trachea
Slides (8)
Ovary Slides (2)
Testis Slides (2)
Duodenum Slides (2)
Spinal Cord (2)
Skin (2)
Hematoxylin
Clearing Agent
Mounting Medium
Coverslips
Slide Staining Dishes (8)
Transfer Pipets (20)
Complete Instructions
CAT. NO.
IND-18 An Introduction to Zoology
PRICE
$115.88
Students prepare and then study sagital
sections of the 72-hour chick embryo in
order to obtain a clear-cut picture of their
developmental anatomies. The picture
above shows a student-prepared slide
Chemical Package Contents
Eosin Y
Isopropanol (2)
Clearing Agent
Mounting Medium
Coverslips
Slide Staining Dishes (8)
Transfer Pipets (20)
Complete Instructions
CAT. NO.
IND-19 An Introduction to
Vertebrate Histology
IND20. AN INTRODUCTION
TO DEVELOPMENTAL
ANATOMY
PRICE
$124.19
72 hr Chick Slides (8) Mounting Medium
Hematoxylin
Coverslips
Eosin Y
Slide Staining Dishes (8)
Isopropanol (2)
Transfer Pipets (20)
Clearing Agent
Complete Instructions
CAT. NO.
PRICE
IND-20 An Introduction to $163.45
Developmental Anatomy
s&!8s7EBSITEWWWMODERNBIOCOMs
CELL BIOLOGY
IND18. AN INTRODUCTION TO
ZOOLOGY
38
IND26. LOCALIZING TUBLIN BY IMMUNOHISTOCHEMISTRY
Microtubules are hollow cylinders made up of polymers of
the protein tubulin. Microtubules are major components
of cilia and flagella, which are tail like projections that
are covered by a plasma membrane and extend outwards
from the cell. Motile cilia are used for locomotion and food
gathering by some protozoa and are found in the lining of the
trachea, where their wave like motion propels mucus, dust
and other foreign matter out of the lungs.
CELL BIOLOGY
In this exercise, students use the powerful technique of
immunohistochemistry to localize tubulin in the esophagus
and trachea. The provided tissue sections are exposed to a
tubulin monoclonal antibody, which binds to the tubulin. The
sections are then incubated with a secondary antibody, which
binds to the first antibody. The second antibody is complexed
with peroxidase, which catalyzes a color producing reaction.
Following the addition of a non-toxic peroxidase substrate,
the peroxidase converts the colorless substrate to a blue
product, thus “staining” the area containing tubulin. As can
be seen in the photographs in panels A-C at the right, the cilia
that line the trachea are stained in a highly selective manner
in keeping with the high concentrations of microtubules that
make up these structures.
IND-26
Localizing Tubulin by Immunohistochemistry
$105.62
(Complete Experiment)
Detailed instructions, 8 unstained trachea/esophagus slides,
clearing agent, mounting medium, coverslips slide staining
dishes, large transfer pipets, small transfer pipets, glass rods, tris
buffer saline, alpha tubulin antibody, peroxidase labeled second
antibody, peroxidase substrate, BSA blocking solution, triton x
100, hematoxylin and eosin.
IND-26AS Tubulin Antibodies and Peroxidase Substrate
$63.19
(Use your own tissue sections)
This kit provides the basic chemicals and antibodies that can be
used to detect alpha tubulin by immunohistochemistry in cells or
tissue sections of your choice. Includes instructions and enough
reagents for the immunohistochemical detection of tubulin in
10-25 slides. Includes 1ml each of alpha tubulin antibody,
peroxidase labeled second antibody, peroxidase substrate
and BSA blocking solution. The IND-26AS does not include
tissue sections, stains, or other solutions.
Microscopes are now
available on page 56.
Panels A-C show the localization of alpha tubulin in the esophagus
and trachea. The pictures were taken at increasing magnification
(40X-1000X). Note that the cilia (indicated by the arrows) that
line the trachea are stained in a highly selective manner. Panel D
shows the esophagus and trachea slides stained with hematoxylin
and eosin.
This exercise was designed for eight groups of students. Five
groups use the above procedure, one group carries out the
procedure without the tubulin antibody (as an important
negative control) and two groups stain their sections with
hematoxylin and eosin as seen in panel D. The analysis is
supplied complete with all of the instructions, chemicals, and
accessories needed to perform perhaps the most important
method used by the cell biologist. Sufficient antibodies,
peroxidase substrate and other reagents are provided so that
between 5-10 additional slides (provided by the instructor)
can be analyzed by the procedure. Distilled water and ethyl
alcohol or isopropyl alcohol are needed but not included.
s-ODERN"IOLOGY)NCss
39
THE OVERVIEW PROGRAMS
The two overview programs were prepared for the beginning college-level student and provide all of the chemicals and
manuals that are needed for 16 students working in pairs to perform an integrated series of experiments on molecular
biology. Each program contains three experiments and each experiment can be completed within a 2-hour laboratory session.
The program manuals contain extensive background information sections and clearly outlined protocols which serve to
acquaint students with molecular biology and with electrophoresis theory and techniques. These Overview Programs provide
stimulating laboratory exercises and a fine introduction to molecular biology for the beginning student.
A Molecular Approach to the
Study of Genetics and Evolution
An Introduction to
Molecular Genetics
Overview Program 1
Overview Program 2
The program contains selected experiments from Standard
Program 1 and stresses the relevance of molecular biology to the
study of human genetics and disease. The experiments provide
an introduction to electrophoresis and to protein composition,
structure, and function in health and disease. The exercises in the
Program are:
This outstanding program provides a solid foundation in molecular
genetics and will prepare your students for advanced collegelevel courses. Your students will investigate such contemporary
topics as DNA electrophoresis, restriction nuclease mapping, and
molecular cloning procedures. Overview 2 contains the following
experiments selected from Standard Program 3:
101.
*102.
*106.
*301.
*302.
*304.
Electrophoretic Separation of Proteins
Genetics and Sickle Cell Anemia
Protein Fingerprinting
*See page 10 for description.
The Length of DNA Molecules
Restriction Nuclease Mapping of DNA
Molecular Cloning (Part A only)
*See page 18 for description.
Price List - Overview Program 2
Price List - Overview Program 1
CAT. NO.
DESCRIPTION
PRICE
O1
The Chemical Package for 16 students working in $107.12
pairs plus one student manual and one instructor
manual. (The student manual may be reproduced for
educational purposes).
01-BK 8 Student Manuals
$48.12
O1-SM Sample Student Manual (29 Pages) plus one
$6.93
instructor manual.
CAT. NO.
DESCRIPTION
PRICE
O2
The Chemical Package for 16 students working in $142.72
pairs plus one student manual and one instructor
manual. (The student manual may be reproduced
for educational purposes.)
Q2-BK 8 Student Manuals
$48.12
O2-SM Sample Student Manual (36 pages) plus one $6.93
instructor manual.
Overview Program 3
This series of three experiments provides an introduction to agarose gel electrophoresis. In the first experiment, students identify
unknown dye molecules by comparing their electrophoretic migration with the migration of known dyes. In the second, they identify dye
molecules that bind to DNA and determine the mechanism. They then study four proteins in experiment 3 and relate differences in their
charges to their migration rates in an electric field. Each protein is a different color so that its progress during the separation can easily
be followed. The experiments were designed for 8 groups of students and the package includes four colorful dye mixtures, four colored
proteins, DNA, agarose, electrophoresis buffer, gel loading transfer pipets and complete instructions. Each experiment can be completed
within a 2-hour laboratory period. The exercises in the program are:
*101.
Electrophoretic Separation of Proteins
*See page 10 for description.
CAT. NO.
DESCRIPTION
PRICE
O3
The Chemical Package for 16 students working in
$54.82
pairs plus 1 Student Manual and 1 Instructor Manual. (The
student manual may be reproduced for educational purposes)
*IND-24.
An Introduction to Electrophoresis
*See page 11 for description.
CAT. NO.
DESCRIPTION
O3-SM Sample Student Manual (29 pages) plus
one instructor manual.
03-BK 8 Student Manuals
s&!8s7EBSITEWWWMODERNBIOCOMs
PRICE
$6.12
$28.16
CELL BIOLOGY
An Introduction to Electrophoresis
40
A Laboratory Course in Innovative Biology (LC6)
This Program is a complete laboratory course for teaching general biology or introductory cell and molecular biology. This program, like
all of our others, is designed for 16 students working in pairs. This course provides essentially all of the chemicals and instructions that are
needed to teach fifteen 3-hour laboratory sessions or twenty-five 1-hour laboratory sessions. The course consists of a series of experiments
that are presented in a comprehensive integrated laboratory manual. In the first two sections of the course, students study topics in protein
biology and biochemistry such as protein structure, function, and isolation. Experiments on enzyme kinetics and cellular metabolism are then
carried out. Students perform a project of their own design in the second section of the course. The projects focus on the characterization of
plant peroxidases. A number of other optional student-designed experiments are outlined in the Instructor manual of the program and basic
reagents are provided in order for the student to carry out their hypothesis driven projects. Experiments on the properties and structure of
DNA are presented in the next section of the course. Here, students perform experiments that deal with genome organization, and specific
gene function. Techniques include DNA electrophoresis, cell fractionation, DNA isolation, restriction nuclease mapping, and basic cloning
procedures. In the final section of the course students study the genetics, biochemistry and molecular biology of hemoglobin.
Features
Suitability – General college-level biology, advanced high school biology or
introductory cell and molecular biology for college sophomores and juniors.
Topics include evolution, protein biochemistry and enzyme action, photosynthesis
and cell respiration, immunology, animal and plant physiology, anatomy and
histology, cell biology, molecular biology and biotechnology, bacteriology,
genetics, molecular genetics and genomics, and human genetic diseases.
Lab Schedule – Fifteen 3 hour lab sessions or twenty-five 1-hour lab sessions.
Cost - $52 per student per student per fifteen 3-hour laboratory sessions, excluding
costs of manuals if the students work in pairs.
Student Designed Experiments - Ideas for over 50 optional student-designed
experiments are outlined in the Instructor manual of the program and basic reagents
are provided in order for the student to carry out their hypothesis driven projects.
Equipment Requirements include horizontal electrophoresis equipment,
microliter dispensers, a microcentrifuge and water bath. A colorimeter is
recommended but not absolutely necessary for parts of two experiments.
Description of the Laboratory Exercises
Proteins and Pigments
1. Electrophoretic and Chromatographic Analysis of
Photosynthetic Pigments from Blue-Green Algae.
Cyanobactera, also known as blue-green algae, obtain their energy by
photosynthesis using sunlight as their energy source. These organisms have
been considered to be the oldest and the most important bacteria on the earth.
It is believed that they were responsible for the initial oxygenation of the earth’s
atmosphere through photosynthesis and it is also felt they were the precursors to
the chloroplasts that are found in true algae and plants. There are two classes of
photosynthetic pigments in Cyanobactera. The first class contains water-soluble
proteins and the major protein is called Phycocyanin, which is blue. The other
classes of photosynthetic pigments that include the carotinoids and chlorophylls
are small molecular weight molecules and are insoluble in water but soluble in
organic solvents such as alcohol. In this laboratory exercise, students isolate and
characterize both groups of pigments. In part A of this exercise, students prepare
a water-soluble extract from blue green algae and show that it contains the single
major protein Phycocyanin by electrophoresis as shown in the gel. They also
determine the charge of this protein by comparing its electrophoretic mobility to
the mobilities of proteins and dyes with known charges. In part B, they prepare
an alcohol extract and analyze the smaller alcohol soluble pigments by thin layer
chromatography in order to identify the chlorophylls and major carotenoid
pigments. The results of this two-part study give students practical hands-on
experience with isolation of components from cells as well as electrophoresis
and thin layer chromatography and introduces them to one of the most important
organisms on the earth.
COURSES
2. Specificity of Albumin Binding
The binding of an enzyme to its substrate is only one example of the many specific
molecular interactions that occur in biological systems. An analogous binding
process occurs with serum albumin, which binds certain small molecular weight
compounds and serves as a carrier molecule for these compounds in blood. In
this exercise, students use an electrophoretic assay to examine the binding of
various dyes to cow albumin. The results of this graphic analysis show that the
binding of dyes to albumin is saturable, specific, compatible, and dependent on
the native structure of the protein.
Enzymes and Metabolism
3. Enzyme Action and Kinetics
This set of experiments was designed to give students a basic understanding
of enzyme kinetics. In the first experiment in this series, students prepare an
extract from wheat germ. They then determined the initial velocity of the reaction
catalyzed by purified acid phosphatase and by the acid phosphatase activity
present in the extract. From these data, they estimate the amount of the enzyme
that is present in the wheat germ. In the second experiment, the student examines
the effects of substrate concentration on the reaction velocity. The results enable
them to determine the Vmax and Km of the enzyme catalyzed reaction.
4. Effects of Temperature on Respiration
Respiration can be viewed as a series of enzyme catalyzed reactions in which
carbohydrates, proteins, and fats are broken down to carbon dioxide and water
with the release of energy. During the process, hydrogen is removed from the
fuel molecules and oxygen is consumed. With this background information,
students measure oxygen consumption and hydrogen liberation in germinating
barley and corn at different temperatures. The program provides eight calibrated
respirometers for measurement of oxygen consumption and the chemicals
required to perform a graphic dye reduction assay.
Cell Biology and Molecular Evolution
5. A Rapid Immunological Method to Study Evolution
Each protein carries in its amino acid sequence information pertaining to its
evolutionary history and origin, and provides clues to the evolutionary history
of the organism in which it is found. Indeed, proteins existing today are in effect
living fossils. This concept is illustrated in this exercise where students examine
the abilities of antibodies against cow gamma globulin to react with gamma
globulins in the sera of cow, goat, sheep, horse, and chicken. The results of the
experiment will enable the student to answer the questions posed in the Student
Guide provided with the exercise. Answers to these questions are given in the
Instructor Guide. The method used for the experiment involves dotting small
quantities of serum onto nitrocellulose and then incubating the nitrocellulose with
an enzyme-linked antibody against cow gamma globulin. Following development
of the nitrocellulose, purple dots appear with intensities that are proportional to
the reactivity of the antibody.
6. Localizing Tubulin by Immunohistochemistry
Microtubules are hollow cylinders made up of polymers of the protein tubulin.
Microtubules are major components of cilia and flagella, which are tail like
projections that are covered by a plasma membrane and extend outwards from
the cell. Motile cilia are used for locomotion and food gathering by some protozoa
and are found in the lining of the trachea, where their wave like motion propels
mucus, dust and other foreign matter out of the lungs. In this exercise, students
use the powerful technique of immunohistochemistry to localize tubulin in the
s-ODERN"IOLOGY)NCss
41
esophagus and trachea. The provided
tissue sections are exposed to a tubulin
monoclonal antibody, which binds to the
tubulin. The sections are then incubated
with a secondary antibody, which binds
to the first antibody. The second antibody
is complexed with peroxidase, which
catalyzes a color producing reaction.
Following the addition of a non-toxic
peroxidase substrate, the peroxidase
converts the colorless substrate to a
blue product, thus “staining” the area
containing tubulin. As can be seen in the
photographs in panels A-C at the right, the
cilia that line the trachea are stained in a highly selective manner in keeping with
the high concentrations of microtubules that make up these structures. In the
course of these studies, students also become familiar with the four basic tissue
types and the structure of tubular organs and identify representative examples of
these features in tissue sections that they prepare.
Student Designed Experiments
7. Characterization of Peroxidase in Plants-Student Designed
Projects
These experiments were designed to actively engage students in exciting
biological research projects of their own design. The projects focus on
peroxidases, which form a large family of related enzymes that are ubiquitous
in plants. Plant peroxidase isoenzymes can be tissue specific, developmentally
regulated and display variable tissue and, high salt and disease resistance defense
reactions and this induction may be
related to the abilities of peroxidases to
strengthen the plant cell wall and to kill
microorganisms. Students begin their
projects with a hypothesis, which is a
statement of an ideal that they will test
in the laboratory. They then test their
hypothesis by carrying out a series of
experiments using the materials provided
with the program and vegetables, intact
plants or roots and stems. They first use
the technique of tissue printing which enables them to localize the peroxidases in
tissue sections. They then quantify peroxidase activity by using a DOT-blot assay
and Spectrophotometric procedures. Students also carry out an electrophoresis
analysis in order to characterize peroxidase isoenzymes in plant extracts that
they prepare. In the final section of the program, students are given detailed
instructions for organizing their data for presentation in a scientific paper. They
are then instructed to write a paper that conforms to the style of a scientific
publication using the detailed steps that are presented in the laboratory manuals.
DNA and the Cell Nucleus
8. Properties of DNA and Cell Fractionation and DNA isolation
A DNA molecule from a single human chromosome is
about 4 cm long and the length of DNA in an individual
is about 200 times the distance from the earth to the
sun. Isolated DNA in a test tube is also a long, stiff
molecule. When alcohol is added to a DNA solution,
the DNA fibers precipitate and can be spooled onto a
glass rod. This feature of DNA is illustrated in the first
part of this lab period. In the second part, students
isolate nuclei from calf thymus tissue. The DNA is then
extracted from the nuclei by a simple procedure that
uses a detergent and alcohol.
Common plasmids are simple DNA molecules, which contain a few genes and
regulatory elements. Most viral genomes are more complex. For example, the
genome of phage lambda contains approximately 50 genes. About 4,000 genes are
present in the E.coli genome while there is approximately 1,000 times more DNA
in the genome of a mammal. This progression in genome complexity is the topic
of this exercise. Here, students compare the electrophoretic patterns of restriction
digests of a plasmid, phage lambda DNA, and cow DNA from thymus and kidney.
Proteins that bind to DNA control the processes of gene regulation and DNA
replication. The electrophoretic mobility band shift assay is a common technique
used to study such specific protein-DNA interactions. In this exercise, students
use this assay to identify dye molecules that bind to DNA and attempt to determine
the mechanism by which these drugs interact with the DNA molecule.
Molecular Genetics Genetics
and Biochemistry of Hemoglobin
11. DNA Cloning and Genotype to Phenotype
This exercise was designed to provide an exciting
introduction to specific gene structure and
function. The students are given four tubes that
are labeled Plasmid A-D. which are identified
in the instructor’s guide. One plasmid (A)
has a functional gene for the enzyme the
ß-galactosidase while tube B contains an inactive
form of this gene because it contains a segment
of foreign DNA. The tube labeled C contains water
while tube D contains the lux operon. Bacteria that
carry this plasmid glow in the dark. In the first part of
the exercise, students analyze restriction digests of the plasmids in order to
determine which plasmid should have a functional ß-galactosidase gene. In
the second part of the exercise, the plasmids are introduced into E.coli by
transformation and the color of the resulting colonies (blue or white) is then
used to assess the functional status of the ß-galactosidase gene. The bacteria
are also viewed in the dark for glowing in order to see which plasmid contains
the lux operon. By comparing the results they identify the plasmids and relate
the genotypes of the plasmids to the phenotypes conferred by the plasmids in
E.coli.
12. Sickle Cell Anemia
Many changes in the structure of hemoglobin have arisen by mutations. About
one person in 100 carries a mutant hemoglobin gene, and these individuals
have abnormal hemoglobin molecules in their blood. One of the most common
abnormal hemoglobins is hemoglobin S, which causes sickle cell anemia.
When the gene for hemoglobin S is inherited from both parents, all of the
hemoglobin in the circulation is hemoglobin S and the individual suffers from
severe anemia. An electrophoretic procedure is used to illustrate the various
types of hemoglobin in the first part of this laboratory exercise. In the second
part, Students use the technique of gel filtration chromatography to isolate
hemoglobin and then to determine its size.
13. Analysis of a Mutant Hemoglobin
Gene
1 2 3 4 5 6 7 8
A mutation is a change in the nucleotide
sequence of DNA, which leads to an
inherited change in an organism. Restriction
endonucleases provide valuable tools for
characterizing mutations at the DNA level. This
principle is illustrated in the exercise where
students digest a normal and a mutant gene
with EcoR1 and Hae III and then analyze the
DNA fragments from each by electrophoresis
as shown in the figure below. The gene is from
rabbit and codes for the ß-globin chain-s of
hemoglobin.
Price List - Laboratory Course 2
CATALOG #
DESCRIPTION
PRICE
LC6
The Chemical Package for 16 students working in pairs
$845
plus one student manual, one instructor manual and a (The student
manual may be reproduced for educational purposes).
LC6SM Sample Student Manual (164 pages) plus one Manual
$37
PDFs
instructor manual
Included
on CD
LC6-BK 8 Student Manuals
$148
s&!8s7EBSITEWWWMODERNBIOCOMs
COURSES
9. Anatomy and Evolution of the Genome
10. Specific Binding of Dyes to DNA
42
A Laboratory Course in College-Level Biology
This program is a complete laboratory course for teaching contemporary biology at the beginning college level. The program
is suitable as a full-semester course that meets once per week and presents an experimental approach to the study of the
biological sciences. The table of contents of the comprehensive program manual is shown on the right. The program is
designed for up to 16 students working in teams of two and provides essentially all of the chemicals, accessories, and
instructions that are needed to teach fifteen 60-90 minute laboratory sessions. Electrophoresis equipment is not required. To
use this program, you will need microscopes, a small centrifuge, ethyl alcohol and acetic acid. The price of the course is about
$29 per student per semester if the equipment listed is available in your teaching laboratory.
As an introduction to the course, students study selected topics in contemporary animal and plant biology including enzyme
action, cell respiration, and evolution at the molecular level. Novel experiments are then presented on mitosis, osmoregulation
and enzyme distribution which serve to help students gain an appreciation for cell structure and function. Protein biological
chemistry forms the next section of the course where the technique of gel filtration chromatography is used to study the structure
and function of biologically important proteins in blood. In
the final section of the program, students are introduced to
Suitability
College-Level General Biology
selected topics on the molecular biology of DNA including
Lab Schedule One 1-2 hour lab session per week for a full
DNA isolation and basic DNA cloning procedures.
FEATURES
The experiments that comprise this course were carefully
selected from Basic Programs 1-4.
Program
Experiment
Description
Basic 1
1, 2, 3
Page 6
Basic 2
1, 2, 3, 5, 6
Page 7
Basic 3
1, 2, 3
Page 8
Basic 4
1, 2, 3, 4, 5
Page 9
Electrophoresis
equipment not
required.
Price List - Laboratory Course 1
COURSES
CAT. NO.
DESCRIPTION
PRICE
LC1
The Chemical Package for 16 students working in pairs plus one
student manual and one instructor manual. (The student manual
may be reproduced for educational purposes.)
$477.16
LC1-BK 8 Student Manuals
$86.35
LC1-SM Sample Student Manual
$21.39
(132 pages) plus one instructor manual.
Manual
PDFs
Cost
semester.
$29 per student per semester excluding costs
of manuals and equipment.
Table of Contents
Introductory Remaks .............................................................................1
Experimental Analysis I: Animal Biology and Evolution
1. The Mammalian Digestive System............................................4
2. Quantifying Amylase Activity ..................................................12
3. Proteases and Factors that Influence Enzyme Activity ............21
4. Evolution of Serum Proteins ..................................................27
Experimental Analysis II: Plant Physiology and Cell Biology
1. Extraction and Analysis of an Enzyme from Wheat ................33
2. Effects of Temperature on Cell Respiration ...........................46
3. Osmolarity and a Cytological Bioassay...................................59
4. Location of an Enzyme in Plant Cells and Tissues ..................67
5. Mitotic Activity and Cell Respiration ......................................74
6. Enzyme Cytochemistry ...........................................................81
Experimental Analysis III: Protein Biological Chemistry and Gel Filtration
Chromatography General Backgrond...................................................88
1. Separating Molecules by Gel Filtration ..................................92
2. The Molecular Weight of Hemoglobin ...................................99
3. Binding Specificity of Serum Albumin .................................104
Experimental Analysis IV: Structure, Isolation, and Function of DNA
General Background ..........................................................................108
1. Properties of DNA................................................................114
2. Cell Fractionation and DNA Isolation...................................119
3. Gene Function and Cloning in Bacteria ...............................126
Included
on CD
s-ODERN"IOLOGY)NCss
43
A Laboratory Course in Cell Biology
This Program is a complete laboratory course for teaching cell biology or introductory molecular biology at the college level.
The program is designed for 16 students working in pairs and provides essentially all of the chemicals and instructions that are
needed to teach sixteen 2-3 hour laboratory sessions. The course consists of 14 experiments carefully selected from Standard
Programs 1, 2, 3, and 7 that are presented in a comprehensive integrated laboratory manual. The Table of Contents of the
manual and the catalog numbers of the experiments provided with the program are shown below. To use this program, you
will need an Accessory Kit, the appropriate electrophoresis equipment, microscopes, and a water bath. A small centrifuge is
required for 2 experiments. The price of the course is about $51 per student per semester if if the equipment listed is available
in your teaching laboratory.
In this laboratory course, your students will be introduced to the molecular biology of the eukaryotic cell. In the first section
of the course, students study topics in protein biology and biochemistry such as protein structure, function, isolation,
molecular evolution, and the detection and molecular basis of human disease. Techniques used for these experiments include
electrophoresis (both native and denaturing), affinity
chromatography, peptide mapping, and the Western blot
Suitability
Eukaryotic Cell Biology or Introductory
procedure. In the second section of the course, students
Molecular Biology for sophomores and juniors.
localize enzymes in plant and animal cells, perform cell
Lab Schedule One 2-3 hour lab session per week for a full
fractionation procedures, and study the properties of a
semester.
specific cell-surface receptor. Experiments on the properties
Cost
$51 per student per semester excluding costs
of manuals and equipment.
and structure of DNA are presented in the final section of
the course. Techniques include DNA electrophoresis and
restriction nuclease mapping.
Table of Contents
FEATURES
The experiments that comprise this course were selected
from four Programs:
Program
Standard 1
Standard 2
Standard 3
Standard 7
Experiment
101, 102, 106
201, 204, 205, 206
301, 302, 303, 306
701, 702, 703
Price List - Laboratory Course 2
Description
Page 10
Page 14
Page 18
Page 33
CAT. NO.
DESCRIPTION
PRICE
LC2
The Chemical Package for 16 students working in pairs plus one
student manual and one instructor manual. (The student manual
may be reproduced for educational purposes.)
$857.76
LC2-BK 8 student manuals.
$93.62
LC2-SM Sample Student Manual (148 pages)
$21.39
plus one instructor manual.
Included
on CD
s&!8s7EBSITEWWWMODERNBIOCOMs
COURSES
Manual
PDFs
Part 1. The Proteins
I. Protein Composition and Structure: A Review of the Basics ..............1
II. Electrophoresis-General Information and Analysis of Native Proteins
Theoretical Aspects ...........................................................................7
Practical Aspects and Procedures. ..................................................11
Experimental Analysis I
1. Electrophoretic Separation Proteins......................................17
2. Genetics and Sickle Cell Anemia............................................20
3. LDH Isoenzymes ....................................................................21
II. Electrophoretic Analysis of Denatured Proteins
General Information........................................................................32
Experimental Analysis II
4. Molecular Weight Determination ..........................................36
5. Peptide Mapping Analysis ......................................................40
6. Protein Evolution and the Western Blo ..................................44
7. Affinity Chromatography ........................................................53
Part 2. The Eucaryotic Cell
General Information........................................................................58
Experimental Analysis III
8. Enzyme Cytochemistry ...........................................................59
9. Analysis of a Membrane Receptor .........................................59
10.The Cell Nucleus ....................................................................86
Part 3. NucleicAcids .............................................................................. I.
Nucleic Acid Structure and Function: A Review of the Basics .............107
II. Electrophoretic Analysis of DNA
General Information......................................................................118
Experimental Analysis IV
11.The Length of DNA Molecules .............................................122
12.Restriction Nuclease Mapping of DNA .................................126
13.Plasmid DNA Structure ........................................................132
14.The Nucleosome Structure of Chromatin ............................136
44
A Laboratory Course in Molecular Biology
This Program is a complete laboratory course for teaching molecular biology at the advanced college level. The program
is designed for up to 16 students working in pairs and provides essentially all of the chemicals and instructions that are
needed to teach seventeen 2-3 hour laboratory sessions. The course consists of experiments carefully selected from Standard
Programs 3, 4, and 5 that are presented in a comprehensive integrated laboratory manual. The Table of Contents of the manual
and the catalog numbers of the experiments provided with the program are shown below. To use this program, you will need
an Accessory Kit, the appropriate electrophoresis equipment, a water bath for DNA hybridization, and a centrifuge that can be
operated at a force of at least 3,000 x g. A bacterial shaker is also strongly recommended. The price of the course is about $39
per student, per semester if the equipment listed is available in your teaching laboratory.
This laboratory course is intended to give students a solid foundation in the molecular biology of DNA and to provide handson-experience with the genetic techniques that form the basis of the biotechnology industry. In the first section of the course,
students use electrophoresis and restriction nuclease mapping to explore important features of prokaryotic and eukaryotic
genomes. DNA hybridization techniques are used in the
second segment of the course where students study molecular
Suitability
Molecular Biology or Molecular Genetics for
evolution at the DNA level and examine a specific sequence in
juniors and seniors.
the eukaryotic genome. In the final section, students prepare
Lab Schedule One 3 hour lab session per week for a full
and characterize a library of recombinant plasmids where
semester.
they receive an appreciation and a detailed understanding of
Cost
$39 per student per semester excluding costs
of manuals and equipment
a variety of state-of-the-art DNA cloning techniques.
FEATURES
The experiments that comprise this course were selected
from three Programs:
Program
Standard 3
Standard 4
Standard 5
Experiments
301, 302, 303, 306
401, 403
Entire Program
Descriptions
Page 18
Page 22
Page 24
Price List - Laboratory Course 3
COURSES
CAT. NO.
DESCRIPTION
PRICE
LC3
The Chemical Package for 16 students working in pairs plus one
student manual and one instructor manual. (The student manual
may be reproduced for educational purposes.)
$658.93
LC3-BK 8 student manuals.
$107.00
LC3-SM Sample Student Manual (118 pages) plus
$21.39
one instructor manual.
Table of Contents
I. Nucleic Acid Structure and Function: A Review of the Basics..........................1
II. Electrophoretic Analysis of DNA
General Information .....................................................................................12
Experimental Analysis I
1. The Length of DNA Molecules............................................................15
2. Restriction Nuclease Mapping of DNA ...............................................20
3. Plasmid DNA Structure ......................................................................25
4. The Nucleosome Structure of Chromatin...........................................30
III. DNA Hybridization Analysis...............................................................................
General Information .....................................................................................48
Experimental Analysis II ...................................................................................
5. Evolution of the Vertebrate Genome ..................................................61
6. Identifying a Specific Sequence in the Mammalian Genome ..............71
IV. Genetic Engineering
General Information .....................................................................................86
Experimental Analysis III
7. Construction of a Recombinant DNA Library .....................................91
8. Transformation of E.coli....................................................................94
9. Isolation of Plasmid DNA .................................................................102
10. Mapping of Plasmid DNA.................................................................113
11. Analysis of Plasmid DNA by Southern Blot Hybridization .................116
Manual
PDFs
Included
on CD
s-ODERN"IOLOGY)NCss
45
A Comprehensive Laboratory Course in Molecular Biology
This Program is a complete laboratory course for teaching molecular biology or biochemistry at the advanced college level.
The program is designed for up to 16 students working in pairs and provides essentially all of the chemicals and instructions
that are needed to teach twenty eight 2-3 hour laboratory sessions. The course consists of experiments carefully selected from
the Standard Programs that are presented in a comprehensive integrated laboratory manual. The Table of Contents of the
manual and the catalog numbers of the experiments provided with the program are shown below. To use this program, you
will need an Accessory Kit, the appropriate electrophoresis equipment, microscopes, a water bath for DNA hybridization, and
a centrifuge that can be operated at a force of at least 3000 x g. A bacterial shaker is also strongly recommended. The price of
the course is about $77 per student per semester if the equipment listed is available in your teaching laboratory.
In this laboratory course, your students will be introduced to many of the concepts and techniques that have revolutionized
the biological sciences during the past two decades. In the first section of the course, students study topics in protein biology
and biochemistry such as protein structure, function, isolation, location in cells, molecular evolution, and the detection
and molecular basis of human disease. Techniques used for these experiments include electrophoresis (both native and
denaturing), affinity chromatography, peptide mapping,
enzyme cytochemistry and the Western blot procedure. In the
second section of the course, students perform experiments
Suitability
Molecular Biology or Biochemistry for juniors
with DNA structure and function. These exercises stress the
and seniors.
organization and complexity of the prokaryotic and eukaryotic
Lab Schedule Two 3-hour lab sessions per week for a fullgenomes, gene function and regulation, and the structure of
semester.
the eukaryotic chromosome. Techniques include restriction
Cost
$77 per student per semester excluding costs
nuclease mapping, Southern blot hybridization procedures,
of manuals and equipment.
and an array of state-of-the-art DNA cloning techniques.
The experiments that comprise this course were selected
Table of Contents
from six programs.
Part 1. The Proteins
I. Protein Composition and Structure: A Review of the Basics .............................................1
II. Electrophoresis-General Information and Analysis of Native Proteins
Program
Experiments
Description
Theoretical Aspects ..........................................................................................................7
Practical Aspects and Procedures ..................................................................................11
Standard 1
101, 102, 106
Page 10
Experimental Analysis I
1. Electrophoretic Separation of Proteins .............................................................17
Standard 2
201, 204, 205, 206
Page 14
2. Genetics and Sickle Cell Anemia........................................................................20
3. LDH Isoenzymes................................................................................................26
Standard 3
301, 302, 303, 306
Page 18
III. Electrophoretic Analysis of Denatured Proteins
General Information ......................................................................................................32
Standard 4
401, 403
Page 22
Experimental Analysis II
4. Molecular Weight Determination ......................................................................36
Standard 5
Entire Program
Page 24
5. Peptide Mapping Analysis..................................................................................40
6. Protein Evolution and the Western Blot.............................................................44
Standard 7
701, 702, 703
Page 33
7. Affinity Chromatography....................................................................................53
FEATURES
CAT. NO
LC4
Price List-Laboratory Course 4
DESCRIPTION
PRICE
The Chemical Package for 16 studentsworking in pairs plus one
student manual and one instructor manual. (The student manual
may be reproduced for educational purposes.)
$1295.16
LC4-BK 8 student manuals.
$127.00
LC4-SM Sample Student Manual (218 pages)
$21.39
plus one instructor manual.
Included
on CD
s&!8s7EBSITEWWWMODERNBIOCOMs
COURSES
Manual
PDFs
Part 2. The Eucaryotic Cell
General Information ......................................................................................................58
Experimental Analysis III
8. Enzyme Cytochemistry.......................................................................................59
9. Analysis of a Membrane Receptor. ....................................................................74
0. The Cell Nucleus ...............................................................................................86
Part 3. Nucleic Acids
I. Nucleic Acid Structure and Function: A Review of the Basics .......................................107
II. Electrophoretic Analysis of DNA
General Information ....................................................................................................118
Experimental Analysis IV
11. The Length of DNA Molecules .........................................................................122
12. Restriction Nuclease Mapping of DNA .............................................................126
13. Plasmid DNA Structure....................................................................................132
14. The Nucleosome Structure of Chromatin ........................................................136
III. DNA Hybridization Analysis ................................................................................................
General Information ....................................................................................................148
Experimental Analysis V .....................................................................................................
15. Evolution of the Vertebrate Genome ................................................................161
16. Identifying a Specific Sequence in the Mammalian Genome ...........................171
Part 4. Genetic Engineering...............................................................................................
General Information ....................................................................................................186
Experimental Analysis VI
17. Construction of a Recombinant DNA Library...................................................191
18. Transformation of E.coli .................................................................................202
19. Isolation of Plasmid DNA ................................................................................207
20. Mapping of Plasmid DNA ................................................................................213
21. Analysis of Plasmid DNA by Southern Blot Hybridization ................................216
46
A Laboratory Course in Cell and Molecular Biology
This Program is a complete laboratory course for teaching cell and molecular biology at the college level. The program is
designed for 16 students working in pairs and provides essentially all of the chemicals and instructions that are needed to teach
sixteen 3 hour laboratory sessions. The course consists of 16 experiments carefully selected from 7 Laboratory Programs that are
presented in a comprehensive integrated laboratory manuals. The Table of Contents of the manual and the catalog numbers of the
experiments provided with the program are shown below. To use this program, you will need an Accessory Kit, the appropriate
electrophoresis equipment, microscopes, a water bath, and a small centrifuge such as tabletop model or microcentrifuge. The
price is about $52 per student per semester if the equipment listed is available in your teaching laboratory.
In this laboratory course, college freshman and sophomore-level students will be introduced to the molecular biology of
the eukaryotic cell. In the first two sections of the course, students study topics in protein biology and biochemistry such
as protein structure, function, molecular evolution, and the detection and molecular basis of human disease. Techniques
used for these experiments include electrophoresis and the Western blot procedure. In the third section of the course,
students localize enzymes in plant and animal cells and study the properties of a specific cell-surface receptor. Students
perform a project of their own design in the fourth section of the course. The projects focus on the characterization of plant
peroxidases. For the projects, assorted vegetables or plants
are needed but not included. Experiments on the properties
and structure of DNA are presented in the final section of
Suitability
Eukaryotic Cell and Molecular Biology for
freshmen and sophomores.
the course. Here, students perform experiments that deal
with genome organization, chromatin structure, and specific
Lab Schedule One 3-hour lab session per week for a full
semester.
gene function. Techniques include DNA electrophoresis, cell
fractionation, DNA isolation, restriction nuclease mapping,
Cost
$52 per student per semester excluding costs
of manuals and equipment.
and basic cloning procedures.
The experiments that comprise this course were selected
from 7 Programs.
Table of Contents
Program
Experiment
Description
Experimental Analysis I: Protein Structure and Evolution
Basic 1
1, 2
Page 6
Protein Composition and Structure: A Review of the Basics............................1
Electrophoresis-General Information and Analysis of Native Proteins
Basic 4
2
Page 9
Theoretical Aspects.........................................................................................7
Standard 1
101, 102
Page 10
Practical Aspects and Procedures .................................................................11
1. Electrophoretic Separation of Proteins ..............................................17
Standard 3
306
Page 18
2. Genetics and Sickle Cell Anemia ........................................................20
Standard 7
701, 702
Page 33
3. Serum Proteins and the Western Press-Blot ......................................26
Experimental Analysis II: Protein Function
Standard 8
801, 803, 804
Page 12
4. Specificity of Albumin Binding...........................................................38
Standard 10 1001, 1002, 1004
Page 20
5. Extraction and Analysis of an Enzyme from Wheat ..............................47
6. Tissue-Specific Isoenzymes in the Cow ................................................59
Others
IND-2, IND-5
Page 34
FEATURES
Price List - Laboratory Course 5
COURSES
CAT. NO.
DESCRIPTION
PRICE
LC5
The Chemical Package for 16 students working in pairs plus one
student manual and one instructor manual. (The student manual
may be reproduced for educational purposes.)
$874.80
LC5-BK 8 student manuals.
$88.68
LC5-SM Sample Student Manual (175 pages)
$21.39
plus one instructor manual.
Manual
PDFs
Experimental Analysis III: Location of Proteins in Cells
7. Enzyme Cytochemistry .........................................................................66
8. Analysis of a Cell Surface Receptor ......................................................81
Experimental Analysis IV: Characterization of Peroxidase in Plants
(Student Designed Projects)
Description ........................................................................................................93
9. Tissue Printing.....................................................................................94
10. Peroxidase Isoenzymes ...................................................................106
Experimental Analysis V: DNA and the Cell Nucleus
Nucleic Acid Structure and Function: A Review of the Basics......................117
Electrophoretic Analysis of DNA
11. Anatomy and Evolution of the Genome............................................130
12. Properties of DNA............................................................................141
13. Analysis of a Genomic Segment .......................................................146
14. Cell Fractionation and DNA Isolation...............................................154
15. Genotype to Phenotype ....................................................................161
16. The Nucleosome Structure of Chromatin ........................................173
Included
on CD
s-ODERN"IOLOGY)NCss