LABORATORY EXPERIMENTS
For
BIO 156
General Biology for Allied Health
Science Department Laboratory
Experiments
COCHISE COLLEGE
SIERRA VISTA
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TABLE OF CONTENTS
Lecture Outlines
Page numbers:
Webstudy login instructions
5
Updating your email address
6
Experiments
1.
Salsa lab/ Scientific Method
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2.
Atoms and molecules (worksheet)
11
3.
Organic molecules (worksheet)
15
4.
Organic molecules biochemical lab tests
21
5.
Microscope lab
29
6.
Cell biology (Prokaryotes/ Eukaryotes)
32
7.
Cell Transport
45
8.
Cell Respiration
51
9.
Overview of metabolism (worksheet)
55
10.
Cellular respiration (Flowchart)
57
11.
Cellular respiration in three acts
59
12.
Mitosis lab
63
13.
Meiosis lab
67
14.
Mitosis/ Meiosis (worksheet)
70
15.
Cancer (Video & worksheet)
75
16.
Genetics Lab 1
77
17.
Genetics Lab 2
79
18.
Nucleic acid and their replication (worksheet)
83
19.
Central dogma and protein synthesis (worksheet)
83
20.
Human DNA extraction
93
21.
Protein Fingerprinting
97
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22.
DNA Unzipped (Zipper Lab)
101
23.
Evolution (Video and worksheet)
125
24.
Evolution Lab 2
127
25..
Antibiotic resistance and natural selection
133
Video and worksheet)
26.
Happy face spiders (Video and worksheet)
135
28.
Human tissues
137
29.
General Orientation to Human Body
141
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1.
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Scientific Method: Salsa Lab
Introduction:
Science is all about describing and explaining the world around us. When scientists make an observation it leads to
questions. Experiments are set up to answer these questions. These questions have to be testable in order to be considered
science. There are many questions that can’t be answered using experiments or observations. These types of questions are
not scientific questions.
Observation:
In this experiment we will all use the same observation. Many people love to eat spicy food, especially in the
Southwestern United States. A person’s reaction to hot and spicy food may be different but at some point, when given
enough spicy food people start to sweat.
Question:
What is a question that we can ask about this observation that can be tested experimentally?
Hypothesis:
Remember a hypothesis is a possible explanation for the natural event. A hypothesis must be falsifiable. Write down
your hypothesis.
Experimental design:
As a group design an experiment that can be used to answer your question and either reject or not reject your hypothesis.
Remember you should have a control and experimental group. Determine what your independent and dependent
variables are. Decide how you will measure your variables.
Record your answers in the lab worksheet.
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Student answer sheet for salsa lab
Name __________________________________________________________________________________________________
1.
What is your question?
2.
What is your hypothesis?
3.
How will you test your hypothesis?
4.
Record your data below.
Time
Student 1
Temp.
Student 2
Temp.
Student 3
Temp.
Student 4
Temp.
Max change in body temp.
5.
Based on all the data collected for the class, fill in the following table:
Mild
Hot
Control 1
Control 2
Mean change in temp.
Standard Error
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6.
Why did we use both mild and spicy salsa in the experiment?
7.
Why do you suppose we took body temperatures under the armpit rather than orally?
8.
Why do you think we have to measure body temperature both before and after you ate the salsa and not just after
(i.e., Why is measuring before and after a better experimental design)?
9.
Which hypothesis was supported? Does this mean it was proven correct? Why or why not?
10. Can you think of any design flaws with this experiment (uncontrolled variables)? If so, what were they?
11. In this experiment which factor was the independent variable(s) and which was the dependent variable(s)?
12. What new questions do you have after doing this experiment?
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Atoms and Molecules
Name ______________________________________________
1. Take a look at the periodic table found in your book and fill in the following table:
Element
Symbol
Atomic Atomic
number mass
Number
of
electrons
Number
of
neutrons
Carbon
Hydrogen
Oxygen
Nitrogen
Helium
Neon
Magnesium
Sodium
Chlorine
Potassium
Phosphorus
Calcium
2. Go to Webstudy and click on the link in this week’s learning unit titled “Chemical Bonds Tutorial.” When the
tutorial loads, be sure to click on and watch each of the following narrations: The tutorial can also be found at
http://bcs.whfreeman.com/thelifewire/content/chp02/02020.html
a. Electron Shells and Chemical Reactivity (click “narrated” to start the tutorial; to get to the next tutorial,
click “options” which will take you back to the main menu).
b. Covalent Bonds
c. Polarity and Hydrogen Bonding
d. Ionic Bonds
3. What makes an atom inert or unable to bond with other atoms?
4. What is a chemical bond?
5. Which atom is the most versatile with respect to bonding and why?
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6. Which type of covalent bond is rare? ____________________________________
7. What is electronegativity?
8. Which atom has a higher electronegativity: oxygen or hydrogen? ______________________
9. What type of bond involves unequal sharing of electrons? _______________________________________
10. If one atom has high electronegativity and another has very low electronegativity, what will happen when they
come in contact with one another?
11. Atoms which have lost or gained electrons are called ________________________________.
a. If an atom LOSES an electron it is called a(n) ________________________.
b. If an atom GAINS an electron it is called a(n) ________________________.
__________________________________________________________________________________________
12. Which of the following contains three elements?
a. H2O
b. CH3NO2
c. CH3OH
d. Na2S
e. Al2O3
13. Which of the following elements is most like calcium?
a. Mg
b. K
c. Sc d. Ne E. Na
14. Bonding. A bond is a connection between two atoms, created when atoms either share electrons to form a
__________________________ bond or when one atom donates an electron to the other atom to form a(n)
____________________________ bond.
a. Examine the water molecule shown in the tutorials you watched. What type of bond occurs between
each hydrogen and the oxygen?_____________________________________
b. Oxygen is a big atom compared to hydrogen. Like a larger planet has greater gravitational pull than a
smaller planet, a bigger atom can have a stronger pull on electrons it shares with a smaller atom. In the
case of water, oxygen draws the electrons it shares with hydrogen closer to itself and farther away from
hydrogen. This creates partial charges within the molecule. Draw a molecule of water then add the
partial charges associated with water (the tutorial might help you). Use the delta () symbol with a + or
– to represent each partial charge.
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c. Is there equal sharing of electrons between atoms in water? ______________
d. Based on your explorations of the water molecule, what specific type of bond does each hydrogen form
with the central oxygen (two words)? _______________________________
15. It’s a rare event to see a water molecule hanging out by itself. Because of the unique kind of bonds found in a
water molecule, a water molecule is __________________ meaning that regions of the molecule are somewhat
more positive, while other regions are somewhat more negative. This property allows one water molecule to
form hydrogen bonds with three other water molecules.
a. Draw a picture below showing four water molecules interacting via hydrogen bonds. Be sure to label
partial charges in each molecule and use a dotted line to represent a hydrogen bond.
b. Is a hydrogen bond a true bond? __________________________________
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Adapted from an assignment by Katie Morrison-Graham, Lane Community College, 2002
Organic Molecules
The questions in italics are ones which you have to complete in class.
The other questions you can answer outside of class if you run out of time.
1.
The players. By definition, all organic molecules contain carbon. Many organic molecules also contain other commonly found
elements such nitrogen, oxygen and hydrogen.
1.1. With the help of your textbook, a molecule model kit and your study partners, complete the chart below to determine the
basic properties of these four important elements.
Element
Symbol
Number of bonds it forms
Carbon
Oxygen
Hydrogen
Nitrogen
1.2. There are a few other elements you’re going to encounter from time to time in this class. Complete the table below to learn a
little bit more about some atoms found throughout your body. Your textbook will help you with this question.
Element
Symbol
Number of bonds it forms
Sodium
Chlorine
Potassium
Phosphorus
Calcium
2.
Bonding. A bond is a connection between two atoms, created when atoms either share electrons to form a
__________________________ bond or when one atom donates an electron to the other atom to form a(n)
____________________________ bond.
2.1. Examine the water molecule in figures from your textbook. What type of bond occurs between each hydrogen
and the oxygen?_____________________________________
2.2. Oxygen is a big atom compared to hydrogen. Like a larger planet has greater gravitational pull than a smaller
planet, a bigger atom can have a stronger pull on electrons it shares with a smaller atom. In the case of water,
oxygen draws the electrons it shares with hydrogen closer to itself and farther away from hydrogen. This creates
partial charges within the molecule. Draw a molecule of water following the ball-and-stick model in figures in
the textbook, then add the partial charges associated with water (figures in book might help you). Use the delta
() symbol with a + or – to represent each partial charge.
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2.3. Is there equal sharing of electrons between atoms in water? ______________
2.4. Based on your explorations of the water molecule, what specific type of bond does each hydrogen form with the
central oxygen (two words)? _______________________________
3.
Molecules interacting. It’s a rare event to see a water molecule hanging out by itself. Because of the unique kind of
bonds found in a water molecule, a water molecule is __________________ meaning that regions of the molecule are
somewhat more positive, while other regions are somewhat more negative. This property allows one water molecule
to form hydrogen bonds with three other water molecules.
3.1. Draw a picture below showing four water molecules interacting via hydrogen bonds. Be sure to label partial
charges in each molecule and use a dotted line to represent a hydrogen bond (figure in the book might help you).
3.2. Is a hydrogen bond a true bond? __________________________________
4.
Get ready to mingle. Water and other polar molecules make for good solutes (dissolvers) because they have the
ability to attract both positively and negatively charged atoms from other molecules.
4.1. Table salt (NaCl) readily dissolves in water. While no true bonds form between Na and water or Cl and water,
hydrogen bonding produces strong attractions between these molecules, splitting apart NaCl to create a
________________________ charged sodium __________ and a _______________________ charged chloride
______________.
4.2. Refer to figure in the book to draw a picture of on NaCl molecule interacting with several water molecules. Label
all molecules and ions appropriately and, as usual, use dotted lines to represent any hydrogen bonds.
4.3. Another common term for ion is (starts with ―e‖) _________________________________________.
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5.
Our friend carbon. Recall that all organic molecules, by definition, contain carbon. Carbon is a dandy molecule
because it forms very stable bonds and is electroneutral. Unlike oxygen, carbon bonds are evenly distributed around
the carbon atom, eliminating the possibility of polar bonds and partial charges.
5.1. Draw a ball-and-stick molecule of methane .
5.2. The chemical formula for water is H20. What is the chemical formula for methane?
5.3. Now imagine if you removed one hydrogen from methane and added a second carbon and its associated
hydrogens. This would create ethane. Draw your completed ethane molecule below (ball and stick model).
5.4. What is the chemical formula for ethane? ____________________________
5.5. Observe your ethane molecule. Are there any polar covalent bonds present? ____________
5.6. Based on your answer would your predict that ethane would or would not dissolve easily in water? ___________
5.7. Now imagine taking off one hydrogen from ethane and adding a hydroxyl group in its place. Draw your
completed ethanol molecule below.
5.8. Ethanol is an alcohol found in drinks such as beer and wine. Examine this molecule. Are there any polar covalent
bonds present? _____________________
5.9. Based on your answer, would you predict that ethanol would or would not dissolve easily in water? ___________
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5.10. Carbon can form bonds with itself and other atoms. Long chains of carbons are common in nature; starch—a
very long chain of glucose molecules—is a good example. Is there any limit (in theory) to the number of bonds
carbon can form with itself? ____________________________
6.
Your basic organic molecule. Organic molecule in the human body fall into four categories.
6.1. What are the four types of organic molecules?
7.
Your pal protein. Proteins form when amino acids bond to each other. Proteins not only lend structure to your body
by helping form things like your bones, but they also protect in the form of immune system antibodies and bloodclotting compounds. Some proteins fall into a special category called catalysts or enzymes. These enzymes, while not
becoming part of a chemical reaction, help speed up a reaction significantly. This comes in handy if you want to
digest your lunch by tomorrow rather than in two weeks!
7.1. Refer to figure 3.4. Draw the general structure of an amino acid.
7.2. An amino acid contains a ____________________ group, an ______________________ group, a hydrogen atom
and another atom or molecule which varies, depending on which amino acid you’re considering. This variable
region is called the ___________________ group.
7.3. The simplest amino acid, ______________________________, contains a ______________ atom in its variable
region.
7.4. How many different amino acids are found in your body? ____________________________
7.4.1. Write down the names and abbreviations for each of these amino acids.
7.5. A ____________ (also called a _____________________ chain) is a long string of amino acids connected together
with covalent bonds and folded into a complex, 3-dimensional shape.
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7.6. What type of reaction joins two amino acids together? ___________________________________________.
7.7. What other types of organic molecules rely on this same reactions to form long chains (there are three)?
____________________________________________________________________________________
7.8. When two amino acids join together, they form a ______________________________ (name of this molecule).
What is the name for the type of bond that forms between amino acids? _______________________________
8.
The sweet stuff. Carbohydrates are also called sugars and have the general chemical formula (CH 2O)n. Based on this
formula, glucose, which contains six carbons (n = 6) would have a specific chemical formula of C 6H12O6. Most sugars
appear as a ring of carbons with oxygen and hydrogens attached.
8.1. Draw and label both glucose and galactose below.
8.2. Look at these two carbohydrates carefully. Both have the chemical formula C 6H12O6, yet they have different
names and different chemical properties. What accounts for this difference if they have the same number of
carbons, hydrogens and oxygens?
8.3. When two carbohydrates bond, they do so through a reaction called ______________________________ (2 words).
This produces a molecule called a _____________________________. A molecule comprising numerous
carbohydrates is called a _____________________________.
8.4. Look at figure in the book. Identify at least three qualities which all carbohydrate molecules share.
8.5. Based on your observations, would you predict carbohydrates are hydrophilic or hydrophobic?
_______________________
8.6. Plants store glucose in a long-chained molecule called __________________________. Your liver stores glucose in
a similar, long-chained molecule, but the molecule has many branches to it. What’s this molecule called?
______________________________ .
9.
It’s what makes food taste so good. Lipids or fats are relatively large molecules that pack a high-calorie wallop (over
9 kilocalories per gram, compared with about 4 kilocalories per gram for either carbohydrates or proteins). While
that’s great if your body’s storing fat to make it through food shortage (imagine how much more you’d weigh if your
body had to store an equivalent amount of energy in the form of carbohydrates!) it’s not so great if you’re consuming
a half gallon of ice cream and trying to watch your weight at the same time.
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9.1. List at least three types of lipids have you learned about.
9.2. What is a primary characteristic of all lipids?
9.3. Based on this characteristic, would you predict they are hydrophilic or hydrophobic? Hint: Think about what
happens when you put a greasy pan into a sink of water without detergent.
9.4. What’s the key difference between a saturated fat and an unsaturated fat ?
10.
Inside information. Nucleotides, when strung together, can create molecules such as DNA and RNA. Both of these
polynucleotides serve as information storage sites in your body. DNA houses instructions we call genes. Genes tell
the cell how to make proteins, but the cell never directly uses DNA to construct a protein—there’s too much risk of
damaging the DNA during protein synthesis. Instead, the cell uses RNA molecules—near-identical copies of the DNA
that forms a particular gene—as a template to make a protein.
10.1. A nucleotide consists of what three parts?
10.2. What four nucleotides are found in DNA?
10.3. What special nucleotide is used as the key energy storage molecule for your body? ____________________
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Organic Molecules Biochemical Lab Tests
Materials (per team of 2 or 3)
1 hot plate
starch solution
12 test tubes
Test tube rack
salt solution
Glucose solution
Distilled water
large beaker
Albumen solution
Reagents: (1 tray per team of 3)
Benedict’s
Silver Nitrate (AgNO3)
Iodine
Sudan IV
Biuret’s (CuSO4 + NaOH)
Benedict’s Test for Simple Sugars
A. Negative Control
1. Pour 5 ml of distilled water into a test tube.
2. Add about 5 drops of Benedict’s to the tube.
3. Place the test tube in a hot water bath.
4. What happened to the color of the solution after heating?
B. Positive Control
1. Pour about 5 ml of glucose solution into a test tube.
2. Add about 5 drops of Benedict’s to the tube.
3. Place the test tube in a hot water bath.
4. What happened to the color of the solution after heating?
Iodine Test for Starch
A. Negative Control
1. Pour about 5 ml of distilled water into a test tube.
2. Add about 5 drops of iodine to the tube.
3. What happened to the color of the solution?
B. Positive Control
1. Pour about 5 ml of starch solution into a test tube.
2. Add about 5 drops of iodine to the tube.
3. What happened to the color of the solution?
Biuret’s Test for Protein
A. Negative Control
1. Pour about 5 ml of distilled water into a test tube.
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2. Add about .5 ml of Biuret’s.
3. What color is the solution?
B. Positive Control
1. Pour about 5 ml of albumen solution into a test tube.
2. Add about .5 ml of Biuret’s.
3. What happened to the solution?
Coagulation Test for Proteins
A. Negative Control
1. Pour about 5 ml of distilled water into a test tube.
2. Heat the test tube in a water bath for about 1 minute.
3. What happened to the solution?
B. Positive Control
1. Pour about 5 ml of albumen into a test tube.
2. Heat the test tube in a water bath for about 1 minute.
3. What happened to the solution?
Sudan IV Test for Fats
A. Negative Control
1. Pour about 5 ml of distilled water into a test tube.
2. Add .5 ml of Sudan IV to the solution.
3. What color is the solution?
B. Positive Control
1. Pour about 5 ml of vegetable oil into a test tube.
2. Add .5 ml of Sudan IV to the solution.
3. What color is the solution?
Silver Nitrate Test for Salts
A. Negative Control
1. Pour about 5 ml of distilled water into a test tube.
2. Add about 5 drops of AgNO3 to the tube.
3. What happened to the color of the liquid?
B. Positive Control
1. Pour about 5 ml of NaCl into a test tube.
2. Add about 5 drops of AgNO3 to the tube.
3. What happened to the color of the liquid?
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Unknown
A. Testing an unknown
1. Obtain a test tube containing an unknown liquid, a 1 cm. square piece of food, or
an egg. If you are using the piece of food move to step 2, if not skip to step 3.
2. Place the 1 cm piece of food in a mortar, add 5 drops of distilled water and crush
the food.
3. Divide the liquid between 6 test tubes. For the egg, use a dropper full per test
tube.
4. Test your unknown using the Benedict’s, Iodine, Biuret’s, Sudan IV and Silver
Nitrate tests.
5. Record your results.
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Organic Molecules
Name/s: __________________________________________
1. Record the results of each of the tests in the following table.
Test
Benedict’s Test
Type
Result
Negative
Control
Positive Control
Iodine Test
Negative
Control
Positive Control
Biuret’s Test
Negative
Control
Positive Control
Coagulation
Negative
Test
Control
Positive Control
Sudan IV
Negative
Control
Positive Control
Silver Nitrate
Negative
Control
Positive Control
2. Before testing your unknown, list your hypothesis of what you think the results will be
for each of the tests.
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3. Record the results of testing your unknown in the following table. Put a positive sign if
the unknown shows a positive result (same result as the positive control) and a negative
sign if it is not.
UNKNOWN
Substance
Tested
Benedict’s
Iodine
Test (+,-)
Test (+,-)
Biuret’s
Test (+,-)
Sudan IV
(+,-)
Silver
Coagulation
Nitrate
Test (+,-)
(+,-)
4. The first five tests are testing for the presence or absence of organic molecules. Using the
results of the tests, what organic molecules are present in your unknown?
5. How does this compare to your hypothesis?
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6. For each organic molecule found in the unknown, list its function in our body and other
examples of foods that contain that organic molecule.
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Microscope lab
Names: ________________________________
Objectives:
1. Identify the parts of the compound microscopes that we will use in this class.
2. Carry and store the microscope properly.
3. Improve your ability to focus the scope using prepared slides (crossed threads, letters), and
examine the given prepared slides.
Obtain a microscope from the cart and carry it back to your desk. Carry the scope with two hands:
one on its arm, and one under the base. Review the parts of the microscope on the scope that you
now have on your desk.
Background:
Microscopes are tools that allow us to extend our senses beyond what we can see with our naked eye.
Before microscopes were invented little was known about the unseen world. Today with compound
light microscopes and even more powerful electron microscopes, scientists are continually learning
more about microbiology and molecular biology, which adds to our knowledge of other levels of
organization. Microscopes are very expensive optical instruments and if given proper care they can
be used for many years.
Parts of the Microscope:
 Stand: The supporting framework of the microscope, which is composed of the base and arm.
o Base: The bottom portion of the microscope and the only part that touches the tabletop.
When carrying the microscope, the upper part of the stand (the arm) is held by one
hand and the other hand should be placed under the base to support the microscope's
weight.
 Stage: The large platform containing an opening near its center. Material to be studied is
mounted on a glass slide and placed on the stage for viewing.
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 Mechanical stage: A mechanism for holding a slide on the stage. The mechanical stage arm
(the silver piece on the stage) will then hold the slide in position. Turning the mechanical
stage knobs at the bottom, right of the stage will move the slide in the x and y directions.
 Illuminator: When turned on with the light switch, it illuminates the field of view.
 Light/power switch: You have a simple on/off switch on the side of your scope. You also have
a rheostat on the side of the scope that adjusts the brightness of the light from the illuminator;
this rheostat on the side of the scope should be set to about 4-5 for normal, bright-field
viewing.
 Condenser: This is a structure located immediately under the stage. Its purpose is to focus the
light coming from the illuminator. A condenser knob under the stage can be turned to focus
the condenser, but the condenser should always be positioned directly under the stage when
you are first examining a specimen; if the image is in focus, but still hard to see, adjust both the
light intensity and this condenser. The condenser has a diaphragm lever attached to it.
Adjusting the diaphragm also will adjust the amount of light coming from the illuminator.
 Eye pieces (Oculars): The lenses of the eye pieces (the oculars) on your microscope magnify
ten times. One of your oculars has a pointer that can be moved by rotating the eye piece. The
pointer is used to show another person a particular object seen in the field of view. You
should focus these double-ocular scopes by closing your left eye and bringing the field into the
proper focus with the focusing knobs on the side of the scope. Then close your right eye and
turn the left ocular until it is in focus. The scope is now focused for the difference between
your right and left eyes. You should also slide the oculars apart so that the distance between
them is the same as the distance between your eyes.
 Focusing knobs: There is one on each side of the lower part of the stand. By turning one of
these knobs, the stage can be moved up and down to bring an object into focus. The coarse
adjustment is the larger knob, and the fine adjustment is the smaller knob. You should ONLY
use the coarse focus when the lowest power objective is in place!
 Revolving nosepiece: Three objectives are attached to this structure. Grip any two objectives
and notice that they can be rotated to allow any one of the objectives to move into viewing
position above the stage. They will "click" into place. If you have trouble seeing one image when
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you look through the oculars, or if the image shape is not circular, it's likely that the revolving nosepiece
is not clicked into place.
o Scanning objective: This lens magnifies four times (4X); it's used to center the object in
the field of view. You will not be drawing at this lowest power, but it serves to initially
orient your slide. This is the only objective with which you should use the coarse focus
knob!
o Low power objective: This lens magnifies ten times (10X) (total magnification = ocular X
objective = 10 X 10 = 100 X total magnification). Focus with the coarse adjustment
knob on scanning power first, then move the nosepiece so that the low power objective
is in place. From this point on, you should never use the coarse focus knob; the fine
focus knob will "fine-tune" the focus that you set up with the scanning objective.
o High power objective: This lens magnifies 40 times (total magnification = 400 X). Use
this objective only after centering an object with the low power objective. You should
only use the fine focus knob when this objective is in place.
o Oil immersion objective: This is the 100X objective. It may be absent in your microscopes,
or some of you may have this as the fourth objective on your microscopes.We will not be using
this lens for the purpose of this class.
IF THE OBJECTIVES OR OCULARS (OR THE SLIDES) ARE DIRTY, CLEAN THEM WITH THE
SPECIAL LENS PAPER. DO NOT CLEAN LENSES WITH TISSUE WIPES, PAPER TOWELS OR
ANY OTHER MATERIAL!
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Student lab sheet for Microscope
1. Label the following diagram and then examine your microscope to identify the corresponding
structures.
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2. What is the magnification written on the ocular lens (eyepiece)?
_____________
3. What is the magnification written on the scanning objective?
_____________
4. What is the magnification written on the low power objective?
_____________
5. What is the magnification written on the high power objective?
_____________
6. The total magnification using the lenses can be determined by multiplying the objective lens
with the ocular lens. What is the total magnification of an item viewed with the:
a. Scanning
______________
b. Low power objective
______________
c. High power objective
______________
7. What is the purpose of the diaphragm?
8. What is the purpose of the stage clips?
9. Look into the eyepiece and twist it left and right. Notice the line inside that moves as
you twist. What do you think this is for?
Using the Microscope
Place the slide (Letter E) on the microscope so that the letter is over the hole in the stage and is right
side-up. Check the diaphragm setting for light source. Click the scanning objective (the smallest
one) into place and use the coarse adjustment knob (the largest one) to focus the slide. If you
cannot get a focus at this point, ask your teacher for help.
Sketch the letter E as it appears under scanning (shortest
objective) power in the circle.
The circle represents the viewing field of the microscope.
Draw your E to scale. (Draw it exactly how it appears in
the microscope, does it take up the whole circle, part of
the circle or some of the circle)
Scanning power
Also note the orientation of the "E", is it right side up or
upside down when viewed thru the lens of the microscope.
33
Have your partner push the slide to the left while you view it through the lens. Which direction does
the E appear to move?
Why would it be important for someone to know that microscopes reverse the image?
Switch the objective to the next level up on the power. (This is the
medium sized objective).
Low Power
Low Power
Use the fine adjustment knob to bring the slide back into focus.
Sketch the "E" as it appears now in the circle. Draw it to scale again.
Now rotate the nosepiece to the HIGH power. Notice the lens should High Power
be very close to the slide. At this point DO NOT use the coarse
adjustment knob. The image should only be focused using the fine
adjustment knob (the little one). Sketch how the E appears now in
the circle.
Slides often get cracked because someone uses the coarse adjustment
while on HIGH power. BE CAREFUL PLEASE!
34
DEPTH PERCEPTION
Obtain a slide with 3 different colored threads on it. View the slide under scanning and low
power.
You should note that while you focus on one color of thread. The other threads become fuzzy. The
microscope can only focus on one area at a time. Sketch the slide below (scanning).
Identify the top, middle, and lower thread colors.
Top thread___________________
Middle thread_________________
Lower thread_________________
35
Choose any 3 specimens from the cart of “common things" such as pond water, euglena, yeast, your
own cheek cells, or any of the permanent slides provided.
Use the circles below to sketch your
specimens under SCANNING/LOW and HIGH power. Label your specimens from the name
written on the slide. Label any structure that you can IDENTIFY.
Specimen 1 _______________________
Scanning Objective
Low Power Objective
High Power Objective
Specimen 2 _______________________
Scanning Objective
Low Power Objective
High Power Objective
Specimen 3 _______________________
Scanning Objective
Low Power Objective
High Power Objective
36
Cell Biology
Prokaryotic and eukaryotic cells
Observation of cells and organelles
In this lab you will be looking at an example of a Prokaryotic cell (Bacillus cereus) and a some
examples of Eukaryotic cells from each of the four kingdoms.
Names and uses for dyes:
Iodine: stains violet/ purple upon contact: Shows the presence of starch
Crystal violet: binds cell wall with starch components
Methylene blue: Binds DNA
Janus Green B: used in histology and to stain mitochondria (changes color depending on amount
oxygen)
Malachite green: used to stain bacteria
Methods
Prokaryotic Cells
Prepare a wet mount of bacterial culture
1. Take a culture tube containing bacteria and re-suspend the cells by rolling the tube between your
palms. The solution should look turbid.
2. Transfer a small drop (the smaller the better) of the culture to the center of a clean microscopic
slide.
3. Observe your sample under the lowest objective, then increase the magnification of your
observations.
4. Place this slide in a 10% bleach container.
Eukaryotic Cells
Kingdom: Protista
Prepare a sample of Euglena for observation
1. Transfer a small drop (the smaller the better) from the container of Euglena to the center of a clean
microscopic slide.
2. Observe the slide under the lowest objective, then increase the magnification of your observations.
3. Label any structures you recognize.
4. Wash and dry your slide and use it for the next procedure.
Kingdom: Fungi
Prepare a yeast sample for observation.
1. Transfer a small drop (the smaller the better) from the beaker containing the yeast culture to the
center of a clean microscopic slide.
2. Observe your sample under the lowest objective, then increase the magnification of your
observations.
37
3. Wash and dry your slide and use it for the next procedure.
Kingdom: Plantae Prepare a slice of potato or carrot tissue for light microscopy. BE CAREFUL NOT
TO CUT YOURSELF.
1. Obtain a small piece of potato or carrot and use a razor blade to shave off as thin a slice as
possible. (i.e. you want to get the white, starchy part of the potato, not the potato skin.) The slice
should be only one or two cells thick, and should appear transparent.
2. Place the tissue slice in the center of your microscope slide, add a drop of water from a dropper
bottle, and then place a cover slip over the preparation.
3. Observe the preparation on the microscope. Remember to start with the lower objective and view
the thinnest edge of your preparation. Draw the cell; be sure to note the magnification of the
image you sketch. Explore the settings of your microscope to determine if you can visualize cell
structure better.
4. Add a drop of iodine to the potato section on your slide. Place a drop of a given solution at one
edge of the cover slip. Touch a lab wipe to the opposite side of the cover slip; the lab wipe will
absorb the water by capillary action and draw the solution over the tissue slice. DO NOT
OVERSTAIN.
5. Observe the preparation again and sketch each sample.
6. Wash and dry your slide and use it for the next procedure.
Prepare an onion slice for light microscopy
1. Obtain a small piece of onion using a sharp blade/ knife. Remove the fleshy “leaves” from a
section and snap the leaf to produce a piece of membranerous tissue. Remove a small piece of
tissue and spread smoothly in a drop of water on a slide. Lower a cover slip gently on top of your
sample to avoid trapping air bubbles.
2. Follow the same procedure for the carrot or potato to observe or stain your onion cells.
Prepare a wet mount of Elodea cells for microscopy
This aquatic plant is useful for observing the typical features of plant cell, which include the cell
wall, chloroplasts, nucleus, streaming cytoplasm and the central vacuole. This leaf is only two cells
thick; you may be able to find differences between the top and bottom surfaces.
1. Cut one leaf from the Elodea plant. Place the leaf in the center of the slide, add a drop of water and
place a cover slip over the mount.
2. Observe the Elodea leaf at various magnifications. Scan the preparation and look for streaming
cytoplasm, which may be more active at the tip of the leaf.
3. Sketch a typical Elodea cell, including the cell wall, chloroplasts, nucleus, and the central vacuole
in your sketch.
4. Wash, and dry the slide and place it on the cart.
Kingdom: Anamalia
Prepare a slide of human cheek cells
Our colorless human cheek cells are transparent and difficult to see, so you will be adding stain to
these cells before viewing them.
38
1. Obtain a clean slide and coverslip and a sterile toothpick.
2. Make a stained wet mount of a human cheek cell: place a drop of methylene blue on a clean
microscope slide. Pick up some of your cheek cells by gently scraping the inside of your cheek
with the side of a sterile toothpick. Transfer the cells to the drop of stain and place a coverslip on
top. (I will show you how to slowly lower a coverslip to prevent air bubbles from entering your
slide).
3. Draw and label a FEW REPRESENTATIVE cheek cells at low AND high powers. (The cells are
too small to see with the scanning objective; remember that you should use this objective only to
orient yourself. Label any visible cellular structures.
The egg as semi-permeable membrane (Demonstration only)
In this demonstration an eggshell filled with distilled water will be floated in a solution of starch and
glucose. The eggshell represents a semi-permeable membrane. The egg will float in the solution for a
time and then we will test to see if the solution in the eggshell has changed or not.
1. What do you think will happen in this experiment. Write down your hypothesis stating what you
think will be the final solution in the eggshell.
2. Record the results of the experiment. How did it compare with your hypothesis?
3. Record your thoughts about what happened in this experiment.
39
40
Cell biology lab sheet
Prokaryotic and eukaryotic cells
Names: _______________________
Always, choose a few, ideal examples and draw them accurately – don’t draw thousands of cells
that all look alike!
Prokaryotic Cells
Record your observations of the bacterial culture.
Eukaryotic Cells
Kingdom: Protista
Kingdom: Fungi
41
Record your observations of the yeast sample.
Kingdom: Plantae
Record your observations of the potato or carrot tissue
Record your observations of an onion.
Record your observations of Elodea cells.
42
Kingdom: Anamalia
Record your observations of human cheek cells.
The egg as semi-permeable membrane
Hypothesis:
Results:
Conclusions:
43
44
Cellular Transport
Osmosis
Make three identical cell models by following these directions.
1. Obtain: Three 250ml beakers and label them 1, 2, and 3
Six pieces of string
Three pieces of pre-soaked dialysis tubing
2. In beaker # 1 add 150 ml of distilled water, bag #1 will be placed into it.
3. In beaker #2 add 150 ml of 5% sucrose solution, bag #2 will be placed into it.
4. In beaker # 3 add 150 ml of 25% sucrose solution, bag #3 will be placed into it.
5. Fold and tie off one end of the dialysis tubing.
6. Open the other end of the dialysis tubing by rolling it between your fingers.
7. Fill each of the bags with 10ml of 5% sucrose solution.
8. Fold and tie off the open end; blot off any excess solution and weigh each bag. Record the initial weights to
the nearest 0.1 gram.
9. Place the bags in their respective beakers at the same time or note the respective start times.
10. After one hour, remove the bags, blot them dry and weigh again. Record the final weight to the nearest 0.1
gram
Simple Diffusion and Brownian Motion
1. Obtain a tiny grain of Congo red dye and place on microscope slide.
2. Add a drop of water on the dye and watch what happens as the dye moves outward from its source.
3. Place cover slip over drop of water and dye and observe under high power. Look for vibrating bubbles. The
goal is to see the dye particles which are large compared to water molecules. The dye particles will appear as
vibrating, clear bubbles. For some reason, this phenomenon of Brownian motion is easy to observe on some
slides and virtually impossible on others, so please share good views with others and be willing to make new
preparations, as this is quickly and easily done. Too much or too little dye does not usually work well.
Demonstration of Diffusion of Substances Through a Semipermeable Membrane
1. Observe Beakers 1 and 2
Beaker one: the dialysis tubing was filled with water and phenolphthalein, then it was
placed in a beaker containing a solution of water and the base, sodium hydroxide
(NaOH). Phenolphthalein is a base indicator that turns pink in basic solutions.
Beaker two: the dialysis tubing was filled with a starch mixture and placed in a beaker
containing a solution of water and iodine. Iodine is a starch indicator that turns dark blue
in the presence of starch.
Osmosis of Decalcified Eggs Placed in Isotonic, Hypertonic, Hypotonic Solutions
1. The instructor will need three student volunteers for this experiment.
2. The initial weight of the eggs will be recorded to the nearest 0.1 gram and be placed in the different
solutions at a recorded time.
3. One hour later, the student volunteers will remove the eggs from the labeled beakers, blot them dry with a
paper towel, and record the final weight to the nearest 0.1 gram.
45
1
Effect of Molecular Weight on Diffusion
Prior to class, grains of the following molecules, potassium permanganate (molecular weight = 158g/mole),
methyl orange (molecular weight = 327g/mole), malachite green (molecular weight = 929g/mole, and
methylene blue (molecular weight = 374g/mole), were placed on separate agar plates at the same time and
allowed to diffuse.
1. Observe the above-described demonstration your instructor has set up.
2. Measure the radius of the halos of color with a ruler for all demos. The halos show how far the substance
has diffused through the agar from its original position.
46
2
Student Lab Write-up for cellular transport
Name: ________________________
Osmosis
a. Complete this table with data from your trial.
Bags with 5%
Initial
Final
Change in Weight
sucrose
Weight
Weight % change = (finalW-initialW)/initialW*100
Bag #1 in
Distilled
water
Bag #2 in 5%
sucrose
Descriptive Term
(solution to bag)
Bag #3 in 25%
sucrose
b. Did water move into, out of, or remain the same in the bags?
Beaker #1
Beaker #2
Beaker #3
c. If a 10% sucrose cell model is placed into a 20% sucrose solution, which direction will water move?
d. Based on your observations of the cell models, is the dialysis tubing permeable to sucrose? Why?
Simple diffusion and Brownian motion
a. Define diffusion.
b. What causes the dye particles to vibrate, that is, why do dead molecules “dance”? What is name given to
this driving force of diffusion?
Diffusion of substances through a selectively permeable membrane.
a. Make a diagram of the initial set-ups for beakers 1 and 2, labeling what was present in each cell model and
in each beaker solution.
b. Describe the appearance of the bag and of the beaker solution in beaker 1 after time has passed.
47
3
c. In beaker 1 which substances were able to pass through the selectively permeable membrane?
d. Describe the appearance of the bag and of the beaker solution in beaker 2 after time has passed.
e. In beaker 2 which substances were able to pass through the selectively permeable membrane?
f. Why are some substances able to pass while others are restricted?
g. Is the transport in these cases considered to be diffusion, osmosis or both? Why?
Osmosis of decalcified chicken eggs.
a. Complete this table with data from your trial.
Egg
Initial
Final
Change in Weight
Weight
Weight % change = (finalW-initialW)/initialW*100
In distilled
water
In 25% sucrose
Descriptive Term
(solution to egg)
In 0.9%
sucrose
b. Did the water move into or out of the eggs in
Beaker 1?
Beaker 2?
Beaker 3?
Which egg gained the most weight? Why?
48
4
Effect of molecular rate on diffusion.
a. Which would you predict would have a faster diffusion rate? Why?
b. Record the radius of the halo of
The potassium permanganate:
The methyl orange:
The malachite green:
The methylene blue:
c. Which substance, the potassium permanganate or the methyl orange, had the largest halo? Is this what you
predicted? Explain
49
5
50
Cellular Respiration
Fermentation
In this activity you will see the action of yeast cells on glucose and distilled water.
1. Obtain 2 Durham tubes and 2 small test tubes per group.
2. Fill one test tube with half yeast solution and half grape juice. Cover the tube with your thumb and shake it
until well mixed. Using a dropper, fill one Durham tube with this mixture, invert it and place it into the test
tube with the grape juice, yeast mixture. No bubble should be in the Durham tube.
3. Fill the second test tube with half yeast solution and half distilled water. Cover the tube with your thumb
and shake it until well mixed. Using a dropper, fill one Durham tube with this mixture, invert it and place it
into the test tube with the distilled water and yeast mixture.
4. Label the tubes with your group name and place them in a test tube rack near a lamp. Both tubes should be
equidistant from the lamp. If they are too close and hot, the yeast will die or be inhibited. If too far, metabolic
levels will be reduced perhaps preventing pronounced results.
5. Allow experiment to incubate for about one hour or longer, then examine and analyze results.
Cellular Respiration and Carbon Dioxide Production: Using Carbon Dioxide as an Indicator of
Cellular Respiration
Carbon dioxide is a byproduct of cellular respiration. Therefore, we can measure the amount of carbon
dioxide as an indicator of cellular respiration. When carbon dioxide combines with water it forms carbonic
acid. Bromthymol blue will be used to detect an increase in carbon dioxide concentration.
Bromthymol blue is a pH indicator, that turns yellow when the pH decreases or becomes more acidic.
In this assignment we will conduct two experiments.
Experiment 1.
In this experiment you will determine if your exhaled breath contains a greater concentration of
carbon dioxide than is present in the atmosphere.
a. Obtain two test tubes and add 3 drops of bromthymol blue into each test tube.
b. Blow into test tube #1 with a straw for 1-3 minutes (careful not to blow too hard and spill).
c. Use a dropper to pump atmospheric air into the second test tube for 2-3 minutes.
d. Record any color changes.
Experiment 2.
In this experiment you will determine if germinating pea seeds produce a greater concentration of carbon
dioxide than is present in the air.
a. Obtain two test tubes and add 3 drops of bromthymol blue to each test tube.
b. Put short segments of glass tubing into the test tubes to make a platform (to keep the peas out of the
solution).
c. In tube #1 place 6-10 germinating peas. In tube #2 do not place anything.
d. Slowly insert a rubber stopper or cotton plug into the tubes, put in a rack, and wait 20
minutes.
e. Record any color changes and results.
51
Demonstration of Cellular Respiration and Heat Production
A few hours earlier two vacuum bottles were set up. Bottle one contains germinating seeds and bottle two
contains only air. Thermometers have been placed in both. Do not open vacuum bottles!
1. Read and record the temperatures of the two bottles.
52
Student Lab Write-up for Cellular respiration
Names: _________________
Fermentation
a. Which Durham tube contained more gas? Why?
b. Did the above experiment require oxygen?
c. What type of gas was captured in the Durham tube?
d. What is the experimental variable in this experiment (remember the experimental variable is the
factor being tested)?
e. What is the dependent variable (remember the dependent variable is what is measured to
determine if the experimental variable has an effect)?
f. How many molecules of ATP are produced when glucose is metabolized during fermentation?
g. Bread is made by mixing flour, water, sugar, and yeast to form dough. Why does the dough rise?
What caused all the concavities in a slice of bread?
h. What “toxic” substance is produced when vertebrates undergo fermentation?
Cellular Respiration and Carbon Dioxide Production
a. Record the results of your experiments in the table below.
Carbon dioxide production in humans and germinating pea seeds
Color of bromthymol blue
CO2 Concentration
after experiment
(circle appropriate)
_______________________________________________________________________
Experiment #1
: Human Expiration
Tube #1: Exhaled air
____________
Increased
No Change
Tube #2: Atmospheric air
____________
Increased
No Change
Experiment #2: Germinating peas
Tube #1: Germinating Peas
__________
Increased
No Change
Tube #2: Atmospheric air
__________
Increased
No Change
b. What is the purpose of tube #2 in experiment 1 and 2?
c. What conclusion can you draw from the results of experiment 1?
d. What conclusion can you draw from the results of experiment 2?
53
Cellular Respiration and Heat Production
a. What is your hypothesis for this experiment?
b. Record the temperatures in each vacuum bottle.
Bottle 1 (germinating peas)_______ 0C Bottle 2 (Air)______0C
c. Do the results support your hypothesis?
d. What conclusion can you draw from the results?
e. Which bottle serves as the control?
54
Overview of Metabolism
Name:____________________
1. Give at least two definitions for respiration.
2. Explain the difference between oxidation and reduction.
3. During reduction, what molecule(s) or atomic particle(s) are usually acquired?
4. Is FAD reduced or oxidized to make FADH2? Explain your answer.
5. What is the definition of metabolism?
55
6. Fill in the following table regarding types of metabolism:
Definition
Dehydration synthesis or
hydrolysis?
Example of this process
Anabolism
Catabolism
7. Distinguish between these three terms:
Aerobic respiration:
Anaerobic respiration:
Fermentation:
56
Cellular Respiration
Name_______________________
begins
with
You may use a term more
than once.
produces a
net gain of
2 ATP
36 ATP
2 NADH
8 NADH
2 FADH2
Electron transport chain
Mitochondrial matrix
Cristae membranes
Cytoplasm
Fermentation
Glycolysis
Glucose
2 pyruvate
Lactic acid
Kreb’s Cycle
which is
shuttled to
which is broken down during
which occurs
in the
which produces
anaerobic
processes
that is used in
such as
aerobic
respiration
which produces
starting with the
which occurs in the
and has a net yield of
used in the
which occurs in the
to make
57
58
Cellular Respiration in Three Acts
Act I: Glycolysis
1. Tell us where this happens.
2. Tell what the purpose of glycolysis is (what is/are the ultimate products?)
3. Identify each actor and tell us who they represent. You should have at least the following players (some
people may have to play more than one role; all people should have a sign to indicate who they are):
a. Glucose
b. 2 ATP (consumed or produced?)
c. Pyruvate
d. Enzymes (don’t worry about identifying individual enzymes, just discuss that they are used to
make the final product)
e. 4 ATP (consumed or produced?)
f. 2 NADH (consumed or produced?)
g. Pyruvate
4. Describe what happens—each actor must ACT OUT their part, not just stand there and smile.
5. Be sure to also address/define the following in your discussion:
a. Is glucose oxidized or reduced to make the final products?
b. Where does the NADH go next?
c. Where does pyruvate go next?
59
Cellular Respiration in Three Acts
Act II: Krebs Cycle
1. Tell us where this happens.
2. Tell what the purpose of the Krebs Cycle is (what is/are the ultimate products?)
3. Identify each actor and tell us who they represent. You should have at least the following players (some
people may have to play more than one role; all people should have a sign to indicate who they are):
a. Pyruvate
b. Acetyl CoA
c. Citrate
d. Isocitrate
e. Alpha-ketoglutarate
f. Succinly CoA
g. Succinate
h. Fumarate
i. Malate
j. Oxalocaetate
k. NADH
l. ATP
m. FADH2
n. CO2 (comes from the break down of what?)
4. Describe what happens—each actor must ACT OUT their part, not just stand there and smile.
5. Be sure to also address/define the following in your discussion:
a. As each intermediate molecule becomes the next molecule, is it being oxidized or reduced?
b. Where does the NADH and FADH2 go next?
c. What are all the final products of the Krebs Cycle?
d. Where does the CO2 come from?
60
Cellular Respiration in Three Acts
Act III: The Electron Transport Chain
1. Tell us where this happens.
2. Tell what the purpose of the Krebs Cycle is (what is/are the ultimate products?)
3. Identify each actor and tell us who they represent. You should have at least the following players (some
people may have to play more than one role; all people should have a sign to indicate who they are):
a. Complex I
b. NAD NADH
c. Complex II
d. FADH FADH2
e. Q
f. Complex III
g. Complex IV
h. ATP Synthase
i. H+
j. Proton gradient (on the other side of the ____?)
k. O2
l. H2O
m. ATP
4. Describe what happens—each actor must ACT OUT their part, not just stand there and smile.
5. Be sure to also address/define the following in your discussion:
a. Where the proton gradient is created?
b. Why the proton gradient is created?
c. Where is oxygen consumed and WHY it is consumed?
d. What does oxygen become after it is consumed?
61
62
Mitosis Lab
Names: _______________________
Introduction
In this lab we will be observing the steps of mitosis under the microscope in the
root tips of plants and the blastula of animals. The process is similar in both
species.
Mitosis of whitefish blastula and onion root tip
Obtain a whitefish blastula slide. Locate cells in each phase of mitosis. Magnify the cells
up to 400x. Draw a picture of each of the cells in the space provided below. If you can
identify them, label the nucleus, spindle fibers and chromosomes with their
centromeres.
Interphase
Prophase
Metaphase
Anaphase
Telophase
Cytokinesis
Obtain an onion (Allium) root tip slide. Locate cells in prophase, anaphase and
telophase. Draw them.
Prophase
Anaphase
Telophase
63
Questions:
Blastula and root tips are embryonic cells. Why would we use embryonic cells to
study mitosis?
Is each cell reliant on the other cells for mitosis or are they independent? Why?
Mitosis Models
As a group, decide which phase of mitosis is shown in the model that was given you.
Give a reason for choosing that phase in the space below. Finally show the teacher what
you decided and why.
Questions:
What percent of the cell cycle consists of interphase?
Does mitotic cell division in eukaryotes yield daughter cells that are identical to or
different from the parent cell? Are the cells different or identical to each other?
What is the difference between cytoplasmic division in plant cells and cytoplasmic
division in animal cells?
64
Comparing Prokaryotic and Eukaryotic division
What are the functions of mitosis in eukaryotic cells?
What is the function of cell division in prokaryotic cells?
How are cell division in prokaryotic and eukaryotic cells similar and different?
65
66
Meiosis: Sexual Reproduction
Names: _______________________
Meiosis
a. Answer the following considering meiosis in humans
1. Number of chromosomes in the parent cell
________
2. Number of chromatids in daughter cells formed by meiosis I
________
3. Number of chromosomes in daughter cells formed by meiosis I ________
4. Number of chromosomes in daughter cells after meiosis II ________
5. Number of haploid cells resulting from meiosis
________
b. Draw a cell with four chromosomes in the stages of mitosis and meiosis listed
below.
Mitosis
Metaphase
Meiosis I
Metaphase
Meiosis II
Metaphase
Anaphase
Anaphase
Anaphase
c. How many chromosomes are found in human cells after mitotic cell division?
d. How many chromosomes are found in human cells after meiotic cell division?
e. Describe the process of meiotic cell division in your own words without using the names of
the phases.
67
f. Meiosis in animals leads to the production of what type of cells? Be specific.
g. What is the number of chromosomes in a human egg and human sperm cell?
h. How many sperm are produced as a result of meiosis? Why this many?
i. How many eggs are produced as a result of meiosis? Why this many?
h. True/False
The zygote, formed by the union of haploid gametes receives:
A random number of chromosomes from each parent.
Equal numbers of chromosomes from each parent.
One member of each chromosome pair from each parent.
Both members of each chromosome pair from each parent.
______
______
______
_____
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5. Mini practical
Examine the three slides or pictures exhibiting mitosis as selected by your instructor. Answer
the questions below.
a. Microscope or picture #1: What stage of mitosis is this cell in? Identify the cell
structure/organelle at the pointer.
b. Microscope or picture #2: What stage of mitosis is this cell in? Identify the cell
structure/organelle at the pointer.
c. Microscope or picture #3: What stage of mitosis is this cell in? Identify the cell
structure/organelle at the pointer
69
Mitosis and Meiosis Worksheet
Name__________________________________
1. Cell division in prokaryotes is called __________________________________________.
2. What are the five stages of mitosis in eukaryotic cells?
3. Define the following terms AND draw and label a picture of each.
a. Sister chromatids
b. Centromere
c. Unduplicated chromosome
d. Duplicated chromosome
70
4. Fill in the table below regarding each hypothetical cell (best to do this one AFTER you attend your f2f class
this week).
Cell
Are the chromosomes
duplicated or
unduplicated?
How many
chromosomes are
present?
How many sister
chromatids are present?
71
5. Match the description with the correct phase
interphase
The chromosomes condense in this phase
prophase/prometaphase
This is the phase where DNA replication occurs
metaphase
The nuclear membrane breaks down in this phase
anaphase
The chromosomes line up at the equator in this phase
telophase
Disassembly of the spindle apparatus occurs in this phase
The sister chromatids are pulled apart in this phase
6. What is a cell plate?
7. Now go to the Webstudy website and click on the assigned week for this worksheet and select the mitosis
assignment hyperlink to be taken to a different website. This website shows cells at an onion root tip (root
tips grow fast) undergoing mitosis. Use the chart below to keep track of each cell as you identify it at the
website. You are going to be shown 36 cells, and the website will help you if you misidentify a cell. Click
NEXT at the bottom of the page to begin tallying cells.
Cell Cycle Stage
Number of cells (use hatch
marks to keep a running count)
Total number of
cells
Interphase
out of 36
out of
Prophase
36
out of
Metaphase
36
out of
Anaphase
36
out of
Telophase
TOTAL
Percentage of cells
in this phase
36
36
36
100%
8. Examine your numbers above. What phase did most of the cells appear to be in?
9. Why do you think most cells tend to be in this phase?
72
10. Identify the name of the phase which matches each description.
Name of Phase
Description
A.
Homologous chromosomes pair up and form tetrad
B.
Spindle fibers move homologous chromosomes to opposite sides
C.
Nuclear membrane reforms, cytoplasm divides, 4 daughter cells
formed
D.
Chromosomes line up along equator, not in homologous pairs
E.
Crossing-over occurs
F.
Chromatids separate
G.
Homologs line up alone equator
H.
Cytoplasm divides, 2 daughter cells are formed
11. Examine the pictures of meiosis below and identify which phase each cell is in by writing the name of the
phase in each box.
From: http://www.biologycorner.com/worksheets/meiosis2.html
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Cancer
Watch the video “cell biology and cancer”
Watch the video at http://www.learner.org/vod/vod_window.html?pid=2042 then answer these questions. Bring
this worksheet to your next f2f class for initials and include it in your portfolio.
Name ______________________________________________
1. Give at least two characteristics of cancerous cells.
2. What are proto-oncogenes?
3. What is point mutation?
4. What is the p53 and what does the functional version of the gene do?
5. What does a mutated version of the p53 gene do or not do?
6. What is a tumor suppressing genes?
7. Cancers are most common in tissues where cell division is common, such as blood cells and cells in the
lining of the lungs or gut. Why is this observation logical?
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76
Genetics Lab 1 Practice Problems
One-trait cross
In humans, brown eyes (B) are dominant over blue (b)*. A brown-eyed man marries a blue-eyed
woman and they have three children, two of whom are brown-eyed and one of whom is blue-eyed.
Draw the Punnett square that illustrates this marriage. What is the man’s genotype? What are the
genotypes of the children?
(* Actually, the situation is complicated by the fact that there is more than one gene involved in eye
color, but for this example, we’ll consider only this one gene.)
In dogs, there is an hereditary deafness caused by a recessive gene, “d.” A kennel owner has a male
dog that she wants to use for breeding purposes if possible. The dog can hear, so the owner knows
his genotype is either DD or Dd. If the dog’s genotype is Dd, the owner does not wish to use him for
breeding so that the deafness gene will not be passed on. This can be tested by breeding the dog to a
deaf female (dd). Draw the Punnett squares to illustrate these two possible crosses. In each case, what
percentage/how many of the offspring would be expected to be hearing? deaf? How could you tell
the genotype of this male dog? Also, using Punnett square(s), show how two hearing dogs could
produce deaf offspring.
Sex-linked traits
In humans, the genes for colorblindness are located on the X chromosome with no corresponding
gene on the Y. It is a recessive allele. If a man and a woman, both with normal vision, marry and have
a colorblind son, draw the Punnett square that illustrates this. If the man dies and the woman
remarries to a colorblind man, draw a Punnett square showing the type(s) of children could be
expected from her second marriage. How many/what percentage of each could be expected?
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Two-trait cross
In Guinea pigs, black hair (B) is dominant over white (b), and rough coat texture (R) is dominant over
smooth (r). Assuming these genes are on separate chromosomes, draw the Punnett square for a cross
between a homozygous black, rough-haired Guinea pig and a white, smooth-haired one. What would
the phenotype(s) of the offspring be? If two of the F1 offspring were crossed, draw the Punnett square
for this cross. Hint: first make a list of the possible gametes, making sure each has exactly one copy of
each of the genes (one allele for each gene). What would the genotype and phenotype ratios be for
the F2 generation?
Baldness in humans is a dominant, sex-influenced trait. This gene is on the autosomes, not the sex
chromosomes, but how it is expressed is influenced by the person’s sex (due to hormones present,
etc.). A man who is BB or Bb will be bald and will be non-bald only if he is bb. A woman will only be
bald if she is BB and non-bald if she is Bb or bb (it’s almost like B is dominant in males and b is
dominant in females). Actually, because of the influence of other sex-related factors, most women
who are BB never become totally bald like men do, but rather, their hair becomes “thin” or sparse. If
two parents are heterozygous for baldness, what are the chances of their children being bald? Use a
Punnett square to illustrate this. Note: because the sex of a person does make a difference in how the
gene is expressed, you need to set this up as a dihybrid cross to account for the sex of the children.
A non-bald man marries a non-bald woman. They have a son and a daughter. If the son becomes
bald, what are the chances that his sister will, too? Use a Punnett square to show this cross.
A woman’s mother is bald, but her father is not. Her older brother is rapidly going bald. She is an
acrobat who hangs by her hair. Should she change her profession before she goes bald, too? Use a
Punnett square to show this.
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Genetics Lab 2
Names: __________________
Monohybrid Crosses with Dominance
a. In corn plants, height is controlled by one gene. Tall (T) is dominant over short (t).
How many alleles controlling plant height would be found in the nucleus of a
corn plant?_________
How many alleles controlling plant height would be found in the nucleus of each
gamete of a corn plant?_________
b. What are the genotypes of the following corn plants?
Homozygous Tall _____ Heterozygous Tall ______
Short______
c. In humans freckles (F) are dominant over non-freckled (f).
Lisa has freckles, but her husband Rick does not. Lisa’s dad has freckles but her mom
does not. What is the chance that their child will have freckles?_______.
What is the phenotypic ratio of the above cross?
What is the genotypic ratio of the above cross?
d. Bob and Mary are both non-freckled. Each had a parent who had freckles. Is it possible for them
to have a kid with freckles? Why or why not?
Personal Genetics Determining Genotype from Phenotype
For each listed trait, record your phenotype, your genotype (to the extent possible) and the ratio of
the class’s genotypes (use given symbols for genotypes).
a) Interlocking Fingers (I = left on top, i = right on top)
Interlock your fingers and observe which thumb you “naturally” place on top. If your left thumb is
on top you have the dominant allele. If right is on top you are homozygous recessive for the trait.
# I_in your group
# ii in your group
# I_in class
# ii in class
b) Hitch-hikers Thumb (J = loose joints, j = tight joints)
Put your thumb up (like Fonzie) and pull it back. If it can go way back, beyond 90 degrees than you
have loose joint ligaments and possess the dominant allele. Otherwise you have tight ligaments and
two recessive alleles for this trait.
# J_in your group
# jj in your group
# J_ in class
# jj in class
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1
c) PTC taste (P = PTC taster, p = PTC non-taster)
Chew the paper provided by your instructor that has been dosed with a harmless chemical,
phenylthiocarbamide (PTC) that some can taste and others cannot. If you are a taster you have a
dominant allele, if you are a non-taster than you are homozygous recessive for the trait.
# P_in your group
# pp in your group
# P_ in class
# pp in class
d) Sodium Benzoate (S = sodium benzoate taster, s = sodium benzoate non-taster)
Chew the paper provided by your instructor that has been dosed with a harmless chemical, sodium
benzoate that some can taste and others cannot. If you are a taster you have a dominant allele, if you
are a non-taster than you are homozygous recessive for the trait.
# S_in your group
# ss in your group
# S_ in class
# ss in class
e) Earlobes (E = free earlobes, e = attached earlobes)
Have a neighbor examine your earlobes. If no part of the lobe hangs below the attachment point,
then you have attached earlobes and have the homozygous recessive condition. Otherwise you have
free earlobes and possess at least one dominant allele.
# E_in your group
# ee in your group
# E_ in class
# ee in class
f) Dimples (D = with dimples, d = without dimples)
Having dimples in one or both cheeks (on your face) is controlled by a dominant gene. Absence of
dimples results from the homozygous recessive condition.
# D_in your group
# dd in your group
# D_ in class
# dd in class
g) Mid-digital Finger hair (H = hair on mid-digit, h = lacking hair on mid-digit)
Examine the back of fingers 3 and 4 (knuckles 1 and 2) for hair. If present, you have a dominant allele
for hair. If absent you are homozygous recessive for this trait.
# H_in your group
# hh in your group
# H_ in class
# hh in class
h) Tongue Rolling (T = able to roll tongue, t = unable to roll tongue)
Stick out your tongue and try to roll it. Rollers have the dominant allele, while non-rollers are
homozygous recessive.
# T_in your group
# tt in your group
# T_ in class
# tt in class
i) Bent Little Finger (L = bent little finger, l = straight little finger)
If the terminal joint of your little finger is angled toward the ring finger, then you are dominant for
this trait. If your pinky is straight, you are homozygous recessive.
# L_in your group
# ll in your group
# L_ in class
# ll in class
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2
j) Sex (XX = female, XY = male)
Well, what can I say? Take a look and mark it down. XX determines female genotype and XY
determines male genotype.
# XX in your group
# XY in your group
# XX in class
# XY in class
k) Freckles (F = having freckles, f = lacking freckles)
Freckles result from a dominant gene. If you lack freckles and always have, you are recessive.
# F_in your group
# ff in your group
# F_ in class
# ff in class
l) Blaze (B = having a blaze, b = lacking a blaze)
A blaze is a lock of hair of a different color. A dominant gene codes for a blaze. If you lack a blaze
you are homozygous recessive for this trait.
# B_in your group
# bb in your group
# B_ in class
# bb in class
m) Widow’s Peak (W = having a widow’s peak, w = lacking a widow’s peak)
If at the center of your hairline you have a v-shape then you have a widow’s peak, the dominant
condition. If your hairline across the forehead is straight you are homozygous recessive for this trait.
# W_in your group
# ww in your group
# W_ in class
# ww in class
n. In every case, if an allele is dominant does the majority of the class possess that allele?
o. If you have a dominant allele for a trait, you do not know whether you are heterozygous for the
trait or if you are homozygous dominant. How could you determine your genotype for this trait?
List two approaches.
Incomplete Dominance and Codominance
Symbols: R=red, r=white
Genotypes and Phenotypes: RR=red, rr=white, Rr=pink
a. What is the genotype of a pink-flowering snapdragon?_______
b. If you crossed two pink-flowering snapdragons what would be the expected
genotypic ratio?_____________________________
phenotypic ratio?____________________________
c. What color of snapdragons would you cross if you wanted all the offspring to be pinkflowering? _________________________.
d. If two parents are heterozygous for sickle-cell anemia, what is the possibility that they
will produce a child with sickle-cell anemia?
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3
Codominance: ABO Blood Types
a. What would be the possible genotypic and phenotypic ratios for the following crosses.
1. IAIB x ii
Genotypic ratio:___________
Phenotypic ratio:___________
A
2. . I i x IB i
Genotypic ratio:___________
Phenotypic ratio:___________
b. True or False
________ A type O child can have two type A parents.
________ A type O child can have one type AB parent.
________ A type B child can have two type AB parents.
________ A type AB child can have one type O parent.
c. Ruby (Type A) is suing Ron (Type AB) for child support for her child (Type B).
Based on blood type is it possible that Ron is the father?_______
What blood type would Ron have to have to prove he was not the father?_____________
d. What are the possible genotypes of a child belonging to parents with AB and O blood
types?
Dihybrid Cross
a. In humans right-handed (R) is dominant over left handed (r), and free earlobes (F) are dominant
over attached earlobes (f). What would be the phenotypic ratio of children born to parents who were
both heterozygous for both right-handedness and free earlobes?
b. What would be the phenotypic ratio of children born to a mom who was heterozygous for both
traits and a dad who was left-handed and had attached earlobes?
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4
Nucleic Acids and Their Replication
Name:_________________________
1. Draw a nucleotide (figure in your textbook) and clearly label its three component parts.
2. Nucleotides found in DNA contain a sugar called dexoyribose, while the sugar group in RNA is called
ribose. Draw the chemical structure for each of these sugars, label each, then CIRCLE the location on
each sugar where they differ from one another.
3. DNA is a polymer. What does polymer mean?
4. What purpose does DNA serve inside a cell?
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5. There are four different type of nucleic acids which may be strung together to make DNA. These four
nucleotides are called _________________________________, _______________________________,
_______________________________ and ______________________. All four of these nucleic acids contain
the same sugar and phosphate groups; where they differ is in their _________________ ______________
(2 words).
6. A fifth nitrogenous base, which is only found in RNA is called _______________________________.
7. Nitrogenous bases in nucleic acids can either be a purine or a pyrimidine. Fill in the chart below
regarding these two general types of molecules.
Chemical structure (draw a picture)
Names of bases of this type
Purine
Pyrimidine
8. In DNA, complementary base pairing dictates that adenine always and only pairs with
_______________________ and ______________________ always and only pairs with
_______________________________.
9. Determine what the complementary strand of DNA would be for the following strand:
T
G
C
G
A
T
G
G
T
C
C
A
10. The process used to join two molecules of nucleic acid together (or any two organic molecules, for that
matter) is called ________________________ ___________________________ (2 words we learned about in
the metabolism lecture, also called a synthesis reaction).
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11. The manner in which nucleic acids join together forms a _________________-_________________
backbone.
12. DNA consists of ___________ polymers of DNA which are joined together by ___________________
bonds between the nitrogenous bases. Because there are two strands in DNA, and because they spiral
around each other, we call DNA a ________________ helix molecule.
13. What defines one end of a DNA molecule as the 5’ end?
a. What defines the other end at the 3’ end?
b. When two strands of DNA are paired together to form a functional molecule, what is interesting
to note about their 5’ and 3’ ends?
14. Synthesize a complementary strand of DNA for this strand:
5’
C
T
A
G
A
C
T
G
A
T
3’
15. DNA synthesis always occurs starting at the _____________ end and moving toward the ______ end.
16. Compared to DNA, RNA is different in a couple ways. Fill in the following table to distinguish the two:
DNA
RNA
Contains these
nucleotides:
Type of sugar?
Double or single
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stranded?
17. In RNA, complementary base pairing dictates that adenine always and only pairs with
_______________________ and ______________________ always and only pairs with
_______________________________.
18. Synthesize a complementary strand of RNA for this strand:
5’
C
U
A
G
A
C
U
G
A
U
3’
19. Which molecule—DNA or RNA—do scientists believe was found in the first life form on Earth? ______
20. Why do scientists believe this molecule was first to form life?
21. DNA synthesis is said to be semiconservative. What does this mean?
22. In order for DNA to be replicated, an enzyme called ______________________ must unwind the two
strands of DNA and break the _________________ bonds between nitrogenous bases. Once open, the
two strands are called the ________________ strand and the _________________ strand. The main
enzyme responsible for synthesizing the new DNA strands is called
__________________________________________ and it requires two “helpers” to get started. These
helpers include an enzyme called ____________ (begins with a “p”), which creates a primer and enough
free-floating nucleotides (called dNTPs or deoxynucleoside triphophates) to make the new DNA.
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23. Synthesis of DNA using the leading strand is a straight forward process. It allows for the formation of a
continuous, new daughter strand which is synthesized starting at the daughter cells ___’ end and
moving away from the ______________________________ fork. The lagging strand, however, cannot be
formed in one, continuous strand because the new daughter strand is formed moving toward the
____________________ fork. Instead, the lagging strand forms via discontinuous replication, which
creates fragments of new DNA called __________ fragments. An enzyme called _____________________
links together the fragments into a continuous strand.
24. An error in copying the DNA nucleotides during synthesis is called a _________________________.
Most of these mistakes are found and removed when DNA polymerase _________________________
(does what?) to the newly formed strand. During the process called _________________ _____________,
a collection of enzymes remove the error and replace it with the correct nucleotide sequence. This
correction process reduces the error rate to only one error for every one billion nucleotides!
25. When UV light strikes DNA at a location where two thymines are side by side, what happens?
26. Xeroderma pigmentosum (XP) is a human disease which is autosomal recessive. It is a genetic mutation
which negatively affects the body’s ability to repair thymine dimers. Without this ability to repair
damage, the individual has a 1000-2000 times great chance of getting skin cancer. Suppose a woman
who is a carrier for XP and marries a normal man. They have two children. One is normal and the other
has XP. What is the genotype of the father?
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88
The Central Dogma and Protein Synthesis
Name___________________________________
1. Draw your own version of the Central Dogma below, being sure to include the terms translation,
transcription and synthesis.
2. Fill in the following chart regarding the Central Dogma:
Transcription
Translation
Goal (outcome)
Required molecules
Location where it
occurs
Post-event
modifications
3. Identify the three main types of RNA and explain the function of each.
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4. What is the genetic code and what is it used for?
5.
According to the wobble rules, the correct amino acid can be added to a growing polypeptide chain
even if the third base in an mRNA codon does not correctly match the corresponding base in a rTNA
anticodon. How do the wobble rules relate to the redundancy of the genetic code?
Synthesize a Protein
6. Below is a region of a gene. Transcribe the gene into a pre-mRNA strand.
DNA
C
mRNA
T
A
T
T
G
C
A
C
C
T
G
A
G
T
C
C
A
7. What is the name of the enzyme which catalyzes this reaction? _________________________________
8. What three things must then happen to the pre-mRNA you just synthesized before it is allowed to exit
the nucleus?
9. Now that you have mature mRNA and it has exited the nucleus and entered the cytosol, it is time to
transcribe the mRNA. You may use the abbreviations for the amino acids found in Figure 15.8 of your
textbook
DNA
C
T
A
T
T
G
C
A
C
C
T
G
A
G
T
C
C
mRNA
Peptide
chain
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A
10. After the polypeptide chain is formed, what or who helps it fold into its final, three-dimensional
structure?
11. Now imagine that a mutation occurred at the first G in the DNA sequence above and the G became a C.
How would this affect the mRNA and the amino acid for that codon?
Old DNA
codon
T T G
Old RNA
codon
Old amino
acid
New DNA
codon
T T C
New mRNA
codon
New amino
acid
12. This would be an example of which type of a point mutation?__________________________
13. Now imagine that a mutation occurred in the second T of the codon below and the T became a G. How
would this affect the mRNA and the amino acid for that codon?
Old DNA
codon
T T G
Old RNA
codon
Old amino
acid
New DNA
codon
T G G
New mRNA
codon
New amino
acid
14. This would be an example of which type of a point mutation?__________________________
15. Now imagine that a mutation occurred in the g of the codon below and the G became a C. How would
this affect the mRNA and the amino acid for that codon?
Old DNA
codon
A T G
Old RNA
codon
Old amino
acid
New DNA
codon
A T C
New mRNA
codon
New amino
acid
16. This would be an example of which type of a point mutation?__________________________
17. Finally, imagine that a mutation occurred in the codon below and an A was inserted between the two
Ts. How would this affect the mRNA and the amino acid for that codon?
Old DNA
codon
T T G
Old RNA
codon
Old amino
acid
New DNA
codon
T AT G
New mRNA
codon
New amino
acid
18. This would be an example of which type of a mutation?__________________________
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92
Human DNA extraction
Introduction: DNA contains the instructions for making you, how you look, what blood type you have, even
your tendency to get some diseases. It is found inside the nucleus in just about every single cell of your body.
In this activity, you will break away the membrane around the cell and its nucleus so that you can see your
very own DNA.
Objective: The objective of this exercise is to extract DNA from human cells (cheek cells).
Materials:
10 ml of .9% salt water in small cup
5 ml of 25% dishwashing soap in 50 ml tube with cap
Test tube rack
1 small cup
1 test tube with 10 ml chilled ethanol
stirring rod
eppendorf tube to save DNA with 0.5ml ethanol
Procedures:
1. Measure 10 ml of salt solution and place it into a small cup.
2. Swish the salt water in your mouth for one minute. Make sure to go the full minute to collect enough
cheek cells.
3. Spit the water into your cup.
4. Pour 5 ml of the dishwashing solution into 50 ml tube.
5. Add the saltwater/cheek cell mixture from the cup to the test tube.
6. Cap the tube and gently rock the tube on its side for 2 to 3 minutes. The detergent will break the cell
membrane releasing the DNA. Be careful not to shake it too hard!
7. Remove the lid and tilt the tube slightly. Pour 10 ml of chilled ethyl alcohol down the side of the tube
so that it will form a layer on top of the soap. DO NOT SHAKE IT!!!
8. Allow the tube to sit in a test tube rack for 1 minute.
9. Place a stirring rod into the first tube and gently wind the DNA onto it. Be careful not to mix the
ethanol and soapy layer.
10. Remove the rod from the tube after winding as much DNA as possible on the stick.
11. If you want to keep this DNA scrape/shake the DNA into the ethanol in the eppendorf tube. Your
DNA should stay solid in this solution.
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Student lab sheet
Human DNA extraction
Names: _______________________________
Answer the following questions:
What does swishing the salt water in your mouth do?
What do you think the extraction solution is? What does it do to the cells?
Why do we add cool ethyl alcohol to the mixture?
Take a look at your tube. What do you see in the top portion of the liquid?
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Protein Fingerprinting
BIG IDEA: Every cell in an organism has the recipe for every single protein in that organism's body, but what
proteins does each cell actually make? During this lab investigation, you will isolate and then separate out the
proteins from different bovine tissues (skeletal muscle, heart muscle, liver, etc.) using protein gel
electrophoresis. This will create a 'protein fingerprint' of different types of cells from a cow so you can examine
whether they make the same or different proteins.
Materials/Equipment Needed
For the class:
• hot water bath or heat block (95°C)
• 2 thermometers
• microwave
• 3% agarose in Tris-Glycine buffer
• Tris-Glycine-SDS buffer
• Coomassie blue stain
• Destain
• Plastic wrap
• 3-4 different bovine tissues
For each student group:
• horizontal gel electrophoresis apparatus, electrodes, power supply
• micropipet with 4 micropipet tips
• 3-4 microfuge tubes with 500 µl sample buffer
• 3-4 empty microfuge tubes
• staining tray
• paper towel or kleenex
Pouring an agarose gel:
1. Get electrophoresis apparatus and make sure there are black stoppers at both ends of the gel tray.
2. Make sure one comb is in place at the negative electrode (black end of the gel).
3. Pour melted agarose into the gel space until the gel fills the shallow tray NOT THE WHOLE BOX.
Let the agarose harden, which should take about 10 minutes. Don’t touch/move your gel until it’s hard.
In the meantime, prepare your protein samples.
Procedure for preparing muscle samples
1. Label each of the 4 sample buffer tubes, one for each type of tissue.
2. Cut a small bit of each tissue (size of half a pencil eraser) and put it into the corresponding tube.
3. Gently shake tubes and let samples sit for 5 minutes.
4. Label 4 empty microfuge tubes, one for each type of tissue.
5. Pipette 1/4 of the liquid (not the tissue!) from your sample tubes to the new tubes.
6. Incubate the tubes in the heat block at 95°C for 5 minutes.
7. The samples are ready to load into the gels. Be sure to keep track of which samples are loaded into
which wells.
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Electrophoresis
1. When the agarose gel is hard, remove the stoppers.
2. Pour Tris-Glycine-SDS buffer over your gel so that is it completely covered plus a little more.
3. Draw a diagram of the gel including the wells. Label which protein samples you are putting into
each well because once the samples are loaded, you will not be able to determine which sample is which.
4. Load as much of each protein sample as you can into its well.
5. Run that gel!! Plug the electrodes into your electrophoresis apparatus (red to red, black to black),
being careful not to bump your gel too much. Plug the power source into an outlet and set the voltage to about
100 V.
6. Let the gel run until the dye migrates about 6-8 cm from the wells (about 20-30 minutes).
7. Turn off the power supply, disconnect the electrodes, and remove the top of the electrophoresis
apparatus.
8. Carefully remove the gel and place it in the staining tray.
9. Pour just enough Coomassie blue stain over the gel to cover it.
10. Cover with plastic wrap and stain for at least 30 minutes. [The gels can stain for longer, but the longer they
stain, the longer they will take to destain.]
11. Remove stain and pour enough destain on the gel to cover it. Tuck a paper towel or some kleenex in the
holder with the gel (it will soak up the stain).
12. Cover with plastic wrap and destain overnight.
13. Put the gel holder with the gel in it on the light box and view your gel. Draw a picture of your gel:
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Student lab sheet for Protein Fingerprinting
Name: ________________
Analysis:
1. below the image of your gel:
2. Compare the protein fingerprints from the different tissues. What can you conclude about what
proteins each type of cell makes?
3. Which tissues had the most similar proteins? Which tissues had the most different proteins? Why do you
think so?
4. Why would proteins from different types of cells look different? Why would they look the same?
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100
DNA Unzipped - Protein Synthesis
Part A
I. In the Nucleus
1. After you receive your egg, open it and remove the contents
2. Examine the drawing inside and read the descriptions.
3. Unzip the zipper. Note that each strand has a 3’ and 5’ end.
4. Pick up the DNA strand written on 3’end – this is the coding strand. Transcribe the DNA bases
into messenger RNA (mRNA) onto the long paper strip provided, following the base-pairing
rules for transcription learned in class.
5. When you are done with transcription, zip up the DNA and replace it in the nucleus. The
mRNA is now ready to leave the nucleus and move to the ribosome. However…
…to avoid bottlenecks at the ribosomes, do the following while still seated:
6. Separate the mRNA into codons by writing a line underneath every three bases on the paper
strip. If you don’t have even groups of three codons (you should have a total of 23 for chain
A or 30 codons for chain B), then you may have missed a DNA base during transcription.
Note: there is a start and stop codon on each strand.
7. Use a codon chart provided to translate the mRNA into amino acid sequence. Write the 3letter abbreviation for each amino acid next to the codon on the paper mRNA.
II. At the Ribosome
8. Carry your mRNA over to a classroom “ribosome”. Cut a string about 10 inches.
9. Following your translated code, construct a polypeptide out of beads. You will need to secure
the first and last bead to prevent them from falling off.
10. Congratulations! You have used the code from a gene to create a protein (or part of one).
Coil and fold your protein to give it a unique 3-D structure. Shortly, you will learn what gene
you decoded from your zipper and what protein you created.
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Checking for Understanding:
a) What does the egg represent?__________________________________________________
b) What does the zipper in the egg represent?_______________________________________
c) What did the long strip of paper represent?________________________________________
d) What is really happening to DNA in a cell when it “unzips”?___________________________
e) When you “copied” the DNA code onto the long strip of paper, what process did this
represent?__________________________________________________________________
f) During transcription, what rules for base pairing did you have to follow? ________________
g) At the ribosome, what did all of the individual beads represent?_______________________
h) When you built your protein out of amino acids by following the coded instructions in RNA, what
process did this represent?_________________________________________________
i) In a real cell, where does transcription occur? ______________Translation?______________
j) Your finished product of beads on a wire represents an example of a __________________.
________________________________
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Part B
Polypeptide hands on activity
Introduction: Students will simulate transcription and translation and learn how proteins are coded from the
DNA nucleotide sequences. Students will understand the role of DNA code, mRNA, ribosomes tRNA in
forming polypeptide chain. They will use the knowledge of DNA base pairing, enzymes, amino acids, proteins
and ribosomes.
Materials:
Two mRNA pieces of paper per group of students
DNA molecules-cut out each group
Have students sort the images: amino acids, cut out, and each separated
Have students cut all the images of: tRNA
Scissors
Scotch tape-one per two students or group
Instructor needs to set up as follows the central dogma activity:
Nucleus: On one corner of the room tape the cut out DNA molecules labeled as “Nucleus” (which is provided
or make up your own) to the board at a height easy read by the students
Cytoplasm: Far from the “Nucleus” now label a region in the room as “Cytoplasm” and spread out all the
amino acids in a tray.
Ribosomes: Tape ribosomes label on each student desk and also leave rolls of scotch tape there.
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Directions: per group of 2 students are going to make two polypeptides.
1. One student in the group is to go the nucleus with the mRNA paper and transcribe the DNA code from
the nucleus to the mRNA molecule. The student must write down the codons onto the space provided,
three letter per underline space. The student returns to their desk and place the code onto the
ribosome.
2. The other partner then translates the code and writes the anti-codon onto the bottom of the tRNA
molecule. The mRNA student looks up the amino acid that tRNA student is supposed to get from the
cytoplasm. (The ribosome is also in the cytoplasm but students are to go to an organized location to get
the amino acids)
3. The tRNA student goes and retrieves the correct amino acids and then brings them back to the
ribosomes and places the tRNA anti-codon at the A site.
4. The tRNA is then moved to the left and the amino acid is left at the P site.
5. The mRNA student looks up the amino acid and the tRNA student transcribes the next codon and
writes down the anti-codon onto a new tRNA molecules. The tRNA student then goes and gets the
next amino acid and brings it back to the ribosomes. The tRNA anti-codon matches with the
corresponding codon.
6. The first amino acid is then taped to the next amino acid. The tape is to represent a peptide bond. The
tRNA is moved to the left and the two amino acids are left at the P site. The amino acids should be
vertical with the first amino acid being farther from the P site. The chain of amino acids should be
perpendicular to the P site and the mRNA.
7. This process repeats until a stop codon is reached. The student brings the stop codon and the amino
acid chain is released, called a polypeptide. Students do not tape the stop codon to the polypeptide.
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Evolution Video “The Evidence for Evolution”
Name: __________________
Give examples of the following evidences of evolution:
1. Fossil Evidence
2. Biogeographical Evidence
3. Morphological/Anatomical Evidence
4. Biochemical Evidence
Define the following terms and give examples of each:
1.
2.
3.
4.
Homologous structures
Analogous structures
Vestigial Structures
natural selection
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Evolution Lab 2
Names: __________________________
1. Introduction
a. Define biological evolution.
b. Are mutations more likely to be harmful or beneficial to an animal in a stable environment? In a changing
environment? Explain why for both.
c. What environmental conditions could increase the rate of evolutionary change?
d. Does gene flow between two populations increase or decrease the probability of them genetically diverging
into two species?
2. Genetic Drift Coin Flipping Experiment
A. With another student at your table, flip a coin 10 times, recording the outcome of each flip by tally in
row 1 of table found below.
B. When finished, record the results of your 10 flips on the chalkboard, and copy the results of 9 other
groups into the table. This will provide a sample size of 100 total flips composed of ten subsets of 10
flips each.
C. Provided a coin is fair, the expected ratio of heads to tails is 1:1 (or 5:5 in a sample of ten). Observe the
results obtained. Suppose the coin flips represent an entire species of 100 animals distributed into ten
populations of ten each, and that each head represents the death of one individual (by a chance event)
beyond the normal mortality rate, while each tail represents one additional viable, offspring beyond the
number usually produced. Compare the impact of these chance events on the population (10 flips) and
species (100 flips) levels.
Heads
Tails
Ratio
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
Total
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a. What conclusions regarding the effects of chance events on the process of evolution may you draw from this
data [hint: compare the effect on the sub-population level (10 flips) and on the entire species level (100 flips)].
b. To further understand the effect of population size on expected events and on evolution, consider human
birth ratio. Human female and male babies are produced in a ratio of about 1:1. Would you expect the ratio of
expected human births to be more closely achieved in a small town hospital or in a large, urban hospital?
Why?
3. Founder effect and genetic bottle neck
a. The smaller the initial population, the more likely that the initial genetic make-up of the group will vary
from the norm for that species. Why?
b. Why might a genetic bottleneck be harmful to a species?
4. Processes that alter gene frequency of populations
a. Define natural selection.
b. Outline the four steps of evolution by natural selection.
c. List five forces that may cause evolution.
5. Fitness
a. Define fitness.
b. Explain the valid statement that "Natural selection acts on the individual, but only populations evolve".
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6. Natural Selection Exercise:
a. To enhance your understanding of natural selection and evolution we will perform a simulation.
Understand and remember what each item represents in the following simulation. We will assume that coat
color of pocket mice is determined by a single pair of genes (alleles).
L = light fur color
l = dark fur color
Cards with two letters written on them will represent each mouse and its genotype for the single trait of fur
color. Black and white beads will represent alternative alleles of the gene for fur color in pocket mice
(remember that an allele is an alternative expression of the same trait or gene, in this case fur color).
White beads = light fur color
Black beads = dark fur color
We will assume a constant population size of 18 mice, determined by the environment.
Because the diploid genotype of each mouse consists of two alleles (2 letters or 2 beads), one from each parent,
and because each mouse only passes on one or the other allele (letter or bead) in each haploid gamete, we must
determine which allele is passed on to each diploid offspring by simulating meiosis. A cup will represent the
gonad that produces the gametes. Since we are only looking at one trait controlled by a pair of alleles, each
one-allele gamete will be represented by one bead. Which of the two beads of the parental diploid genotype is
passed on will be determined by randomly drawing (close your eyes or hide the cup when drawing) one bead
from the cup. Beyond illustrating the action of natural selection, this simulation will also reinforce the
principles of meiosis and Mendelian inheritance.
Simulation 1: No Selective Pressure
1. To begin, obtain 18 cards and make sure that the genotype frequency of the 18 mouse population is as
follows: 6 LL, 6 Ll, 6 ll. This will be generation 1. Understand how allele frequencies are calculated from
genotypes.
2. Place the 18 cards face down, mix them up and randomly draw to make 9 pairs of cards (9 pairs of parents).
This disregards male and female, which is OK for this simulation as we assume that each pair consists of one
male and one female. Each pair of parents will produce exactly two offspring (replacement level
reproduction) and then the parents will die.
3. Produce two offspring for each pair by simulating meiosis for each parent and combining the winning
gametes (beads) to make the genotype of a mouse offspring. This must be done twice to make two offspring.
Suppose that you randomly drew cards with Ll and ll as a pair of parents. First you would select the gamete
passed on by Ll by placing a white and a black bead in a cup and randomly drawing one bead. Then you
would do the same for the other parent, ll, by placing two black beads into the dish and drawing one (or by
simply calculating that ll can only pass on a black bead or l allele). Combining the two drawn beads would
provide the genotype of one offspring (either Ll or ll depending on whether a black or white bead was drawn
for the first parent). Record the offspring’s genotype in simulation 1 table. You would repeat the process for
the second offspring. After producing two offspring, those two parents die. Repeat for all nine pairs of
parents producing 18 genotypes of offspring.
4. Assume that each pair of parents of generation one dies after producing two offspring each. The mice of
generation 2 are the offspring from generation 1.
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5. Make 18 new cards with the genotypes of the parents for next generation and repeat steps 2-4 until the 7th
generation of mice are produced. This means when you finish generation 1 and have 18 offspring genotypes
marked in generation 2 (the offspring of generation 1 and future parents of generation 3) that you must remark
the cards to reflect the genotypes of generation 2, etc. If you do not change the cards, you are simply repeating
generation 1 over and over.
Simulation 1: No Selective Pressure
Generation
Parental Genotypes
LL
Ll
1
2
6
6
ll
Allele Frequencies
L
l
6
18
18
3
4
5
6
7
**b. Compare the genotype and allele frequencies of generations 1 and 7. Are they the same or do they differ?
It is predicted by the Hardy-Weinberg Principle that they should be similar unless mutation, gene flow,
selective breeding, genetic drift or natural selection acts on the population. Which of these five forces may
have affected this first simulation? Explain. Read and understand simulation 2 (assignment 7) before
answering.
7. Natural Selection Exercise:
Suppose that a flow of dark lava invaded an environment, producing dark soils and making light-furred mice
very evident to predators. This simulation will address such a scenario.
Simulation 2: Selection Pressure Against Light Fur
1. To begin, obtain 18 cards and make sure that the genotype frequency of the 18 mouse population is as
follows: 6 LL, 6 Ll, 6 ll. This will be generation 1. Understand how allele frequencies are calculated from
parental genotypes.
2. Place the 18 cards face down, mix them up and randomly draw to make nine pairs of cards (nine pairs of
parents). This disregards male and female, which is OK for this simulation.
3. Produce four offspring for each pair of parents by simulating meiosis for each parent and combining the
drawn gametes (beads). Write down the genotypes of all four offspring on scratch paper. Only two of the four
offspring will survive because natural selection will eliminate two of the offspring (preferentially killing light
furred mice). The survivors are determined by the following "survival rule": ll>Ll>LL. Which means that the
"L" allele is selected against and "l" allele is selected for. Simply eliminate the two offspring with more "L" and
less "l". Record your two surviving genotypes after elimination of the two less fit offspring in the simulation
table. Repeat for all nine pairs of parents producing 18 genotypes of offspring.
4. Assume that the parents of generation 1 die after producing four offspring each. Only two of four offspring
live, determined by survival rule in step 3 and environmental constraints. The parent mice of generation 2 will
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be the two surviving offspring from each generation 1 mouse. Successively repeat steps 1-4 until the 7th
generation of mice are produced.
5. Make 18 new cards with the genotypes of the parents for next generation and repeat steps 2-4 until the 7th
generation of mice are produced.
a.
Simulation 2: Selective Pressure Against Light Fur
Generation
Parental Genotypes
Allele Frequencies
LL
Ll
ll
L
l
1
2
6
6
6
18
18
3
4
5
6
7
b. What is unrealistic about the elimination rule, that is how do genetics and natural selection really work?
**c. Compare the genotype and allele frequencies of generations 1 and 7. Are they the same or do they differ?
It is expected that they should be similar unless mutation, gene flow, selective breeding, genetic drift or natural
selection acts on the population. Which of these forces affected this simulation? Explain, comparing the
results of simulations 1 and 2.
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Antibiotic Resistance and Natural Selection
Name: ________________________________________
Watch the video at http://www.pbs.org/wgbh/evolution/library/11/2/real/e_s_6.html then answer these questions.
Bring this worksheet to your next f2f class for initials and include it in your portfolio.
1. Why is the Russian prison system considered to be "ground zero" in the fight against TB?
2. What is responsible for the evolution of TB strains that are resistant to multiple drugs?
3. How does the misuse of antibiotics affect the evolution of disease-causing bacteria? Use the theory
of natural selection to explain the growing resistance to antibiotics.
4. Why should we care about a resistant strain of TB in Russia?
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An Example of Evolution: Happy Face Spiders
Name ______________________________________________
Based on information at http://evolution.berkeley.edu/evolibrary/article/happyface_01 (accessed June 23, 2006)
1.
Happy face spiders are endemic to Hawaii. What does endemic mean?
2.
Happy face spiders can vary considerably in appearance, yet they are all considered to be members of the
same species.
a. What is the definition of species?
b.
Why are all the individuals in the picture above considered to be members of the same species?
Provide at least two reasons.
3.
What is morphology?
4.
Yellow happy spiders account for about two-thirds of the population of happy spiders on all islands,
while the other one-third of happy face spiders varies in color. Researchers first thought dispersal would
describe why this ratio was constant across islands. What evidence and what about the evidence led
researchers to reject the dispersal hypothesis?
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5.
6.
Which of the Hawaiian islands is the youngest? ____________________________________
What does this have to do with the happy face spiders?
7.
Why did the researches reject the idea that sexual selection (females being attracted to males of a
particular color morph) accounted for the 2:1 ratio in happy face spiders?
8.
9.
Who eats happy face spiders?______________________________________________________
What explanation did the researchers eventually provide to explain the 2:1 ratio?
10. Is their explanation (hypothesis) correct? Explain your answer.
11. Can (or have) the researches prove(n) their hypothesis? Explain your answer.
12. Why is genetic variation important for natural selection?
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Human Tissues
Names: _____________________
1. Epithelial tissue
a. Draw the following epithelial tissues as you seem them under 100X total
magnification.
simple squamous epithelium
simple cuboidal epithelium
simple columnar epithelium
stratified squamous epithelium
ciliated psuedostratified epithelium
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b. Match each phase with the correct epithelial type. More than one answer may
apply.
1. simple squamous epithelium
2. simple cuboidal epithelium
3. simple columnar epithelium
______Found in kidney tubules
_____Lines blood vessels
______Makes up the skin
4. stratified squamous epithelium
5. ciliated psuedostratified epithelium.
_____Lines upper respiratory tract
______Has goblet cells
2. Connective Tissue
a. Draw the following connective tissues as you seem them under 100X total
magnification.
Loose connective tissue
Adipose Tissue
Dense connective tissue
Hyaline cartilage
Osseous (Bone) tissue
Vascular tissue (blood)
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b. Match each phrase with the correct connective tissue type.
4. vascular tissue
1. loose connective tissue
2. dense connective tissue
5. adipose tissue
3. osseous tissue
6. hyaline cartilage
______Stores fat
______Has a hard matrix
______Has a liquid matrix
______Has a soft matrix with all 3 fiber
______Makes up fetal skeleton
types
______Makes up tendons and ligaments
3. Muscle Tissue
a. Draw the following connective tissues as you seem them under 100X total
magnification.
Skeletal muscle tissue
Cardiac muscle tissue
Smooth muscle tissue
b. Match each phrase with the correct muscle type. More than one answer may apply.
2. cardiac muscle
3. smooth muscle
1. skeletal muscle
______Voluntary
______Has a hard matrix
______Involuntary
______Striations
______Spindle shaped cells
______Branching cells
______Intercalated discs
______No striations
4. NervousTissue
a. Draw a neuron as you see it under 100X total magnification.
Neuron
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5. Review of general tissue types
a. Match each phrase with the correct tissue type. More than one answer may apply.
2. Connective
3. Muscle tissue
1. Epithelial tissue
tissue
4. Nervous tissue
______receive and conduct electrical impulses
______its cells may contain striations
______contracts to produce movement
______provides protection and support
______contains more extracellular matrix than cells
______ cells are called neurons
______its cells form glands
______lines body cavities
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General Orientation to Human Anatomy
Anatomical position
The subject stands upright, feet flat on the floor, arms at the sides and palms facing forward.
Anatomical right and left refer to the subject's right or left.
Directional Terms
Directional terms are used to explain where one body structure is in relation to another. For example,
we say the head is superior to the neck.
List of directional terms and definitions
1. Superior (cephalic or cranial): toward the head or above other structures
2. Inferior (caudal): away from the head or below other structures
3. Anterior (ventral): toward the front or belly surface
4. Posterior (dorsal): toward the back of the body
5. Medial: toward the midline
6. Lateral: away from the midline
7. Intermediate: between two structures
8. Proximal: nearer to the attachment of extremity to the trunk
9. Distal: farther from the attachment of extremity to the trunk
10. Superficial: towards the body surface
11. Deep: away from the body surface
Regional Terms
Regional terms are used to describe body regions and landmarks. Many of these words have Latin or
Greek origins. Parts of these terms are often used when naming muscles and bones.
List of regional terms and definitions
1. Cephalic: pertaining to the head
2. Occipital: pertaining to the back of the head
3. Nasal: pertaining to the nose.
4. Buccal: pertaining to the cheek
5. Orbital: pertaining to the eye
6. Oral: pertaining to the mouth
7. Cephalic: pertaining to the head
8. Occipital: pertaining to the back of the head
9. Cervical: pertaining to the neck
10. Thoracic: pertaining to the chest bone
11. Sternal: pertaining to the breast
12. Abdominal: pertaining to the abdomen
13. Umbilical: pertaining to the navel region
14. Coxal: pertaining to the hip
15. Sacral: pertaining to the area between the hips
16. Pubic: pertaining to the genital region
17. Pelvic: pertaining to the pelvis
18. Scapular: pertaining to shoulder blade region
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19. Deltoid: pertaining to curve of shoulder formed by deltoid muscle
20. Vertebral: pertaining to the spine
21. Lumbar: pertaining to lower back
22. Inguinal: pertaining to the groin
23. Acromial: pertaining to the shoulder
24. Axillary: pertaining to arm pit
25. Brachial: pertaining to the arm
26. Carpal: pertaining to the wrist
27. Digital: pertaining to the fingers
28. Antecubital: pertaining to the anterior surface of the elbow
29. Olecranal: pertaining to posterior surface of the elbow
30. Gluteal: pertaining to the buttocks
31. Femoral: pertaining to the thigh
32. Popliteal: pertaining to the posterior knee area
33. Patellar: pertaining to the anterior surface of knee
34. Fibular: pertaining to the lateral side of the leg
35 Crural: pertaining to the leg
36. Sural: pertaining to the calf
37. Calcaneal: pertaining to the heel of foot
38. Plantar: pertaining to sole of foot
39. Tarsal: pertaining to the ankle
Body Planes
Body planes are imaginary, flat surfaces used to divide the body for reference.
Body planes
1. Sagittal plane: divides the body or organ into right and left halves.
Midsagittal plane: special case of sagittal plane, that divides body or organ into equal right and left
halves.
2 Frontal (coronal) plane: divides the body or organ into anterior and posterior portions.
3. Transverse (horizontal) plane: divides the body or organ into superior and inferior halves.
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General orientation to human anatomy
Names: _____________________
Lab report questions
1. Anatomical position
a. Describe anatomical position.
2. Directional terms
a. Use the following directional terms to complete the sentences.
Superior, inferior, anterior, posterior, medial, lateral, intermediate, proximal, distal, superficial, deep.
Terms are only used once.
1. The lips are _______________to the teeth.
2. The eyes are ____________ to the nose.
3. The wrist is ______________ to the elbow.
4. The head is ____________ to the chest.
5. The muscles are ____________ to the skin.
6. The elbow is ____________ to the wrist.
7. The skin is _______________ to the bones.
8. The stomach is _________________ to the heart.
9. The nose is ___________ to the eyes.
10. The teeth _____________ to the lips.
11. The heart is ____________ to the arms.
3. Regional terms
a. Write in regional terms that match the descriptions.
__________________1. Navel
__________________16. Ankle
__________________2. Posterior surface of knee
__________________17. Lower back
__________________3. Wrist
__________________18. Mouth
__________________4. Armpit
__________________19. Leg
__________________5. Hip
__________________20. Head
__________________6. Curve of shoulder
__________________21. Heel
__________________7. Arm
__________________22. Calf
__________________8. Neck
__________________23. Thigh
__________________9. Fingers
__________________24. Cheek
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_________________10. Eye area
__________________11. Back of the head
__________________12. Knee (anterior)
__________________13. Genital area
__________________14. Sole of foot
__________________15. Chest
__________________25. Spine
__________________26. Nose
__________________27. Groin
__________________28. Buttocks
__________________29. Abdomen
__________________30. Shoulder
4 Body Planes
a. Write in the body planes that match the descriptions.
________________1. Divides body or organ into right and left halves.
________________2. Divides the body or organ into anterior and posterior halves.
________________3. Divides the body or organ into superior and inferior sections.
________________4. Divides the body into equal right and left halves.
5. Organ Systems
a. Match the organ system to the organs and functions below.
a. Integumentary
e. Endocrine
i. Digestive
b. Skeletal
f. Cardiovascular
j. Urinary
c. Muscular
g. Lymphatic
k. Reproductive
d. Nervous
h. Respiratory
______1. Testes, prostate, penis
_____12. Transports blood
______2. Lymph nodes, tonsils
_____13. Produce offspring
______3. Skin, hair, nails
_____14.Eliminates wastes
______4. Nasal cavity, trachea, lungs
_____15.Gas exchange
______5. Heart, blood vessels
_____16. Coordinates body activity
______6. Stomach, intestines, esophagus
_____17. Protection, vitamin D synthesis
______7. Pineal, pituitary and adrenal glands _____18. Picks up excess fluids
______8. Brain, spinal cord, sensory receptors _____19. Breaks down food
______9. Cartilage, bones, joints
_____20. Movement
______10. Skeletal muscles
_____21.Protects and supports organs
______11. Kidney, bladder, urethra
_____22. Secretes hormones
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6. Body Cavities
a. Match the following body cavities with their descriptions. Each is only used once.
a. Dorsal body cavity
e. Thoracic cavity
b. Ventral body cavity
f. Abdominal cavity
c. Cranial cavity
g. Pelvic cavity
d. Spinal cavity
_______1. Divided into the thoracic, abdominal and pelvic cavities
______2. Contains the brain.
______3. Contains the reproductive organs
______4. Divided into the cranial and spinal cavities
______5. Contains heart and lung
______6. Contains the spinal cord
______7. Contains the stomach, spleen, liver.
b. Label the diagram
1.__________________
2. __________________
3. __________________
4. __________________
5. __________________
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