Wood to Wheels- Inquiry Lesson Plan
Lesson Introduction

Title: Genetic Engineering with Bio-Rad pGLO

Subject/ target grade: 9-12

Duration: 3-4 class periods (55 minutes)

Setting: classroom, computer lab, science lab
 Learning Objectives:
Students will be able to:
 Describe the necessary steps involved in genetic transformations and how
scientists select for transformed (transgenic) organisms.
and treatment.
 Demonstrate lab sterile technique.
 Apply the concept of genetic engineering to other aspects of human health
 Explain the structure and function of bacterial plasmids as they relate to biotechnology
and gene therapy
 Use the RAFT writing strategy to express knowledge and opinions regarding the ethical
dilemma of cloning
 Michigan Content Expectations:
Inquiry Expectations
 1.1 Scientific Inquiry
o 1.1A Generate new questions that can be investigated in the laboratory or field.
o 1.1E Describe a reason for a given conclusion using evidence from an investigation.
o 1.1I Distinguish between scientific explanations that are regarded as current
scientific consensus and the emerging questions that active researchers investigate.
 1.2 Scientific Reflection and Social Implications
o 1.2A Critique whether or not specific questions can be answered through scientific
investigations.
o 1.2B Identify and critique arguments about personal or societal issues based on
scientific evidence.
Cell Structure and Function
 B2.5i: Relate cell parts/organelles to their function
DNA/RNA and Protein Synthesis
 B4.1B: Explain that information is passed by genes that code for DNA, these
genes produce proteins
 B4.2B: Recognize that every species has its own characteristic DNA sequence
 B4.2C: describe the structure and function of DNA
 B4.2D: Predict the consequences that changes in DNA composition of a
particular gene may have on an organism
 B4.2f: Demonstrate how genetic information in DNA molecules provide
instructions for assembling proteins

B4.2g: Describe the process of replication, transcription and translation and how
they relate to each other in molecular biology
 B4.4c: Explain how mutations in the DNA sequence of a gene may be silent or
result in phenotypic change in an organisms and in its offspring
Mendelian and Molecular Genetics:
 B4.2h: Recognize the genetic engineering techniques provide great potential and
responsibilities
 B4.4a: Describe how inserting, deleting or substituting DNA segments can alter
a gene
 Lesson Overview:
This lesson is centered on the Bio-Rad pGLO Bacterial Transformation Kit. Students will examine
how a gene and associated trait from one organism can be isolated, transferred to another organism and
replicated. Students will also use the RAFT writing strategy to assess knowledge about the ethical
dilemma of cloning.
Lesson Core
 The Guiding Question:
What applications can be derived from bacterial transformation?
 Materials and Equipment Needed:
 Bio Rad pGLO transformation kit. ***This kit contains enough supplies for 2 classes of 32
students each. Additional equipment not included is listed below.
 Incubator (37 ° C)
 Hot water bath (42° C) Using DI water will prevent mineral deposits.
 2-4 U-V/ black lights (green fluorescent protein cannot be observed without this light)
 Ice and small Styrofoam cups
 Sharpie markers or wax pencils – 1 per group
 Masking tape
 Buckets or other containers with 10% bleach solution
 Safety precautions:
The E. coli used in this lab in non-pathogenic, but should be treated as such in the lab.
All participants should wear goggles and gloves. All lab supplies, except paper products,
should be placed in a 10% bleach solution and soaked overnight. Teacher can then place items
in a double plastic bag and place in the trash. Agar should NOT be disposed of down the sink drain as
it will solidify and block the drain.

Advanced Preparation: For Bio-Rad pGLO Bacterial Transformation Kit
Teacher Preparation
Step 1 Read through the transformation manual
Make a copy of Student Manual and
Quick Guide for each student
When
Immediately
Time
1 hr
Step 2
Prepare nutrient agar plates
3–7 days prior
1 hr
Step 3
Prepare starter plates, aliquot solutions
36–48 hr prior
30 min
Step 4
Organize student workstations
Immediately prior
10 min
Bacterial cultures need to be started 24 hours before the lab and should be no older than 36 hours. LB
and other plates can be made up to 1 week before the lab and stored in the refrigerator. Instead of
disposing of equipment after bleaching, many items can be washed in hot, soapy water and used for
other labs throughout the year. Autoclaving is another option.
 Background Information for Teachers: (from Bio Rad pGLO Transformation Kit)
In this lab, students will perform a procedure known as genetic transformation. Genetic
transformation occurs when a cell takes up and expresses a new piece of genetic material—DNA.
This new genetic information often provides the organism with a new trait, which is identifiable
after transformation. Genetic transformation literally means change caused by genes and involves
the insertion of one or more gene(s) into an organism in order to change the organism’s traits.
Genetic transformation is used in many areas of biotechnology. In agriculture, genes coding for
traits such as frost, pest, or drought resistance can be genetically transformed into plants. In
bioremediation, bacteria can be genetically transformed with genes enabling them to digest oil
spills. In medicine, diseases caused by defective genes are beginning to be treated by gene therapy;
that is, by genetically transforming a sick person’s cells with healthy copies of the defective gene
that causes their disease.
Genes can be cut out of human, animal, or plant DNA and placed inside bacteria. For example, a healthy
human gene for the hormone insulin can be put into bacteria. Under the right conditions, these bacteria can
make authentic human insulin. This insulin can then be used to treat patients with the genetic disease,
diabetes, because their insulin genes do not function normally.
With the pGLO transformation kit, students use a simple procedure to transform bacteria with a
gene that codes for Green Fluorescent Protein (GFP). The real-life source of this gene is the
bioluminescent jellyfish Aequorea victoria, and GFP causes the jellyfish to fluoresce and glow in
the dark. Following the transformation procedure, the bacteria express their newly acquired jellyfish
gene and produce the fluorescent protein, which causes them to glow a brilliant green color under
ultraviolet light.
In this activity, students will learn about the process of moving genes from one organism to
another with the aid of a plasmid. In addition to one large chromosome, bacteria naturally contain
one or more small circular pieces of DNA called plasmids. Plasmid DNA usually contains genes for
one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer
plasmids back and forth, allowing them to share these beneficial genes. This natural mechanism
allows bacteria to adapt to new environments. The recent occurrence of bacterial resistance to
antibiotics is due to the transmission of plasmids.
 Engage:
This video clip from the 1931 movie version of is a great discussion starter on life and the idea of using
parts from other life forms to create a new life or new traits.
"Its Alive" Frankenstein Clip http://www.youtube.com/watch?v=8H3dFh6GA-A
This is a classic scene. Almost everybody has heard the famous line,” it’s alive." This scene sets up the
outlandish idea that you can take dead parts from a once living thing, put them together, shock it, and
create a new life. Nobody could actually think that it would be possible. However, in less than 100
years from the making of this movie science has unlock the secret of life, in the form of DNA, genes
and genetics.
 Building on prior knowledge:
After the clip, prompt the class with question about the reality of such an idea. Discuss the
difference between physical traits vs. genetic traits. How can a non-living thing bring about life? What
is the value of all the parts to make a whole organism, why not just half a man? There are a thousand
questions to ask and the students always have something to say or plenty of questions, which leads to
more discussion.
 Pre-teaching:
The units preceding the this unit would include mitosis / cell reproduction, a comprehensive unit on
DNA and protein synthesis. Much of the prior knowledge needed to link and understand concepts
would be taught in these preceding units. The PowerPoint from lesson plan 1 will also contain
background information important to this lesson
 Explore:
Bio-Rad pGLO Bacterial Transformation Lab (use kit)
The pGLO lab includes a detailed materials list, step by step and day by day instructions along with a
Teachers Guide with answers and a Student Manual with picture lab flow chart and questions. There is
little teacher prep, the students prep the lab the first day and spend the next 2 days on the experiment.
 Explain:
Students will complete Bio-Rad pGLO Bacterial Transformations Lab Data and Analysis with Review
Questions. Answers will be shared and discussed as a class. (Possible answers are included in BioRad pGlo Bacterial Transformation Lab manual. If time permits, students could instead create a lab
report with their findings.
(**Optional Lab Report Template and Rubric)
Lab Report Template
Title:
* A brief, concise, yet descriptive title
Statement of the Problem:
* What question(s) are you trying to answer? * Include any preliminary observations or background
information about the subject
Hypothesis:
* Write a possible solution for the problem.
* Make sure this possible solution is a complete
sentence.
* Make sure the statement is testable.
Materials:
* Make a list of ALL items used in the lab.
Procedure:
* Write a paragraph (complete sentences), which explains what you did in the lab. * Your procedure
should be written so that anyone else could repeat the experiment.
Results (Data):
* This section should include any data tables, observations, or additional notes you make during the
lab. * You may attach a separate sheet(s) if necessary.
* All tables, graphs and charts should be
labeled appropriately
Conclusions:
* Accept or reject your hypothesis.
* EXPLAIN why you accepted or rejected your hypothesis using
data from the lab.
* Include a summary of the data - averages, highest, lowest..etc to help the reader
understand your results
* List one thing you learned and describe how it applies to a real-life
situation. *Discuss possible errors that could have occurred in the collection of the data
(experimental errors) *How could you improve your procedure ? * After completing this lab, what
other questions would you like to test?”
Lab Report Rubric
Introduction
(4 pts)
(3 pts)
(2 pts)
(1 pt)
1. Includes the question to be
answered by the lab
2. states hypothesis that is based on
research and/or sound reasoning
3. title is relevant.
4. Hypothesis (prediction) is testable.
One of the
"excellent"
conditions is not
met
Two of the
"excellent"
conditions is not
met
Three of the
"excellent"
conditions is not
met
A description or
step-by-step list
of how the
experiment was
performed
Description
unclear, couldn't
be repeated
Results are
present, though
too disorganized
or poorly
recorded to make
sense of
Methods
Results and data are clearly recorded,
organized so it is easy for the reader
to see trends. All appropriate labels
are included
Results are clear
and labeled, trends
are not obvious,
Results are
unclear, missing
labels, trends are
not obvious at all
Analysis
The data and observations are
analyzed accurately, trends are noted,
enough data was taken to establish
conclusion
Analysis somewhat
lacking in insight,
enough data,
though additional
data would be
more powerful
Analysis lacking
in insight, not
enough data was
gathered to
establish trends,
OR analysis does
not follow data
Analysis poor,
not enough data,
inaccurate
analysis
Conclusions
1. Summarizes the essential data used
to draw conclusions
2. Conclusions follow data (not wild
guesses or leaps of logic),
3. Discusses applications of
experiment ("real world" connections)
4. Hypothesis is rejected or accepted
based on the data.
One of the
"excellent"
conditions is not
met
Two of the
"excellent"
conditions is not
met
Three of the
"excellent"
conditions is not
met
Neat, organized
with headings,
few
spelling/grammar
errors
Somewhat
lacking in
organization,
multiple
spelling/grammar
errors, not neat
Results
(data)
Format

(0)
Elaboration: (*** I have my own RAFT write up for this cloning activity- but as it is at home,
I included this one that mine is modified from)
RAFTS writing strategy- taken from:
http://udhscff.wikispaces.com/RAFT+Strategy
For this assignment I want you to think about what we have learned this semester
about the science behind stem cell/somatic cell research, cloning and the various
social and moral dilemmas that are associated with this controversial topic. Is this a
way that science could harm us, or help us? What are the repercussions? What does
cloning mean to mankind? You may take a position for or against, either way, just be
sure to defend your stance citing scientific facts and examples as well as note the
opposing side’s argument. Review the R.A.F.T. sheet attached, then, you must decide
who you would like to be when presenting your position. For example, you can be the stem cell
writing to the President or a clone writing to Congress. You may choose any writing style you would
like.
Guidelines
Review R.A.F.T.S. sheet to review what choices you would like to use to make your writing
piece original.
Minimum of two pages and a maximum of four, this will also depend on your format. Write
enough to defend your point.
Writing piece must be typed, Times New Roman, 12 point font and 1.5 spacing
Be Creative with this assignment if the format supports pictures and you think they are
compelling enough to add to your piece please do so.
Creative Writing Assignment – To Clone or Not To Clone, That is the Question
Role
Audience
Format
Topic
Strong Verbs
Who are you
going to be?
Who are you
writing to?
What type of
document are
you preparing?
What is the
subject
and situation?
What is the intent
Of your writing
piece?
You
A Somatic
Cell
The
President
Speech
Letter
A Researcher
looking to
develop this
technology to
take over the
world
A Human
Clone
An Animal
Clone
You may choose any
of the above roles
To:
Persuade
Congress
News Article
A Researcher
looking to
develop this
technology to
further
medicine
Subject:
Cloning
A Research
Board
Offering
Grants
The
Multiple
Sclerosis
Association
Persuasive
Essay
Undercover
Report
Debate
Any Other
Clone
Situation: Use
all the scientific
facts that you
know or
research to
support your
position either
for or against
cloning.
Support your
position with
examples.
Assess
Compare
Urge
Analyze
Contemplate
Defend
The General
Public
You may choose
any of the above
audiences
You may choose any of
the above formats
Review the above
verbs and decide
how you would
like to proceed
with your writing
- Example
Dear My Beloved Body,
All I am is a tiny somatic cell, why does everybody make such a big deal about me? I am
asking you, my body, because it wasn’t that long ago that you tried to kill me. All I wanted was to go
pay a quick visit with a few of my neighbor somatic cells. I didn’t know there was already a party
going on over there, and, I must admit, it got extremely crowded. The next thing I can remember is we
were getting chemotherapy. Do you know that I lost some very dear friends and family during that
internal war? Not to mention I was sick for a long time. Now, here we are a few years later, and you
are trying to harvest me to make a new pancreas for yourself. You can’t seem to make up your mind!
First, you try to kill me, now you want to extract me and command me to do you a favor. What’s with
this therapeutic cloning, I thought they just cloned sheep?
My understanding of therapeutic cloning is they take me, one of your closest friends and
remove my nucleus. My nucleus is special to me, I really am not pleased to have it removed! Then, to
top it off, you want to stick my nucleus with an egg cell that has also had its nucleus removed! To
think, you want me to move in with someone I haven’t even met! To top that off, we both then need to
move to what I like to refer to as another planet – a uterus! This is where the union would produce an
offspring with my exact genetic makeup. I believe there are wonderful reasons to allow these types of
procedures. For example, there would no longer be a need for people having transplants to go on
immunosuppressant drugs. It would also allow patients to live a life without the potentially serious
side-effects of immunosuppresant drugs. More importantly, the ability to clonally derive organs would
alleviate the current shortage of transplants and would possibly reduce waiting times for translants to
become available. Conversley, if you were looking just to create another you, that would scare me.
Morally and ethically, there is only one you. You have been with me from birth and if this is what I
need to do to extend your life and make it better for you, then, for you my beloved body, I will do it.
Take care of your new organ, I wouldn’t want to have to do this again.
Love,
Your little somatic cell
 Evaluate:
Students make posters presenting their RAFT’s. Students present their RAFT’s to the class, the
following rubric is used to assess
RAFT Rubric
4
Information, details in
RAFT always
accurate. Properly
reflects information,
ideas, and themes
related to the subject.
Accuracy
3
Provides accurate
information in
RAFT but could use
more support.
2
1
Provides
information in
RAFT that has
some inaccuracies
or omissions.
Provides
information in
RAFT that is
incomplete and/or
inaccurate.
Perspective
RAFT maintains clear, Explains how
consistent point of
character would feel
view, tone, and ideas
about the event(s).
relevant to role
played; ideas and
information always
tied to role and
audience.
Shows little
insight into how
character would
feel or act during
the event(s).
Does not
accurately
develop
characters,
thoughts or
reactions to the
event(s).
Focus
RAFT stays on topic,
never drifts from
required form or type;
details and
information are
included that are
pertinent only to
developed purpose.
Uses class time
appropriately to
research the era and
create well-written
stories.
Spends most of the
RAFT discussing
issues on topic, but
occasionally strays
from the focus.
Spends some time
discussing issues
off topic.
Spends most of
RAFT on issues
that do not
directly deal with
the RAFT chosen.
Seldom needs to be
reminded to get
back on task.
Uses library and
computer time to
do work for other
classes and/or chat
with friends or
lounge on
couches.
Treats research
time as an open
period to be seen
chatting with
friends and
hanging out on the
couches.
Essay contains few to
no fragments, run-on
sentences; rare errors
or mechanical
mistakes; writing is
fluent.
Essay contains some
fragments, run-ons
or other errors;
occasional mistakes;
writing is generally
clear.
Essay contains
several sentence
errors and
mechanical
mistakes that may
interfere with
ideas and clarity
of ideas in
writing.
Essay contains
mechanical
mistakes; is
marred by
numerous errors.
Class Time
Mechanics
A+ (20)
below)
A (19)
A- (18)
B+ (17)
B (16)
B- (15)
C+ (14)
C (13)
D (12)
F (11 and

Lesson Closure:
Questions that the teachers might ask to bring the big scientific idea of the lesson.
Questions that the teacher might ask to assess mastery of the learning objectives.
Lesson Extension

Assessment Options:
1. Bio-Rad pGLO Bacterial Transformation Data and Analaysis with Review Questions
2. Lab Report- assess with Rubric
3. RAFT writing strategy- assess with rubric

Additional Resources:
Cloning RAFT taken from: http://udhscff.wikispaces.com/RAFT+Strategy
It’s Alive! You tube clip: http://www.youtube.com/watch?v=8H3dFh6GA-A