DESIGN YOUR OWN GEL ELECTROPHORESIS/RESTRICTION ENZYME LAB
Goal: With your table partners, design an experiment that investigates some
aspect of gel electrophoresis or restriction enzyme digestion.
Evaluation: Your lab grade for this project is based on the following –
-
your experimental design (How well does your experimental design
test your experimental question?) 4 pts
your scientific work (How well do you run your experiment?) 4 pts
your lab book (How well do you record your experiment?) 5 pts
Powerpoint presentation to document all aspects of the project
(question, experimental design, results, analysis). More details to
follow. 12 pts
Potential variables for Gel Electrophoresis:
Potential variables for Restriction Enzyme digest:
Supply List:
(Buffer followed by restriction enzymes in italics)
Buffer C – RsaI
Buffer D – SalI
Location(s) of RE sites on lambda DNA can be found at
Buffer E – BamHI, HindIII, SspI
www.fermentas.com/techinfo/nucleicacids/siteslambda.htm
Buffer H - EcoRI
Buffer K - ScaI
Multicore Buffer
Agarose
BSA
TBE
37oC Incubator
Refrigerator
Heat Block
Loading Dye
Lambda DNA
Gel Electrophoresis equipment (casting tray, gel boxes, power supply) 1 per group
NOTE #1: Your experimental design may not include:
- more than two gel boxes or casting trays on any given day
- more than four restriction enzyme digests
NOTE #2: I will stain any DNA gels with ethidium bromide and take Polaroid
pictures of stained gels. However, I will not stop gels for you unless you tell me the
precise time that you want them to be stopped. Alternatively, you may come in to
stop the gels yourself.
NOTE #3: Outside of any Polaroid pictures of DNA gels, what other data will you
collect? What pictures do you want to show everyone? I STRONGLY ADVISE YOU
to take multiple digital pictures of your experiment to include in your presentation.
Each of the following steps must be approved by me. Don’t proceed without
the approval initials first!
STEP #1 – DEVELOP YOUR EXPERIMENTAL QUESTION (First come, first
served) Ex. How does temperature affect how quickly DNA moves through a gel?
Our experimental question –
Seal of Approval
STEP #2 – DEVELOP YOUR EXPERIMENTAL DESIGN How are you going to test
your question? If you are setting up a restriction digest, you must set up a chart to
show me what is going into each tube and how much is going to be used. If you don’t
ask for it, I may not necessarily have it ready for you and this may impact your
ability to get started ASAP.
Seal of Approval
LAB BOOK
Your lab book should contain the following elements:
1. Title (included in Table of Contents as well)
2. Experimental question
3. Explanation of experimental question (why did you choose this question?
What did you think would happen? Include a diagram of anticipated results.)
4. Results – clearly labeled
5. Analysis – thinking about your experimental question, what do you conclude
based on your results? What experiment would you suggest doing next?
POWERPOINT PRESENTATION
1. Explain what your experimental question. What was it? Why did you choose
that one? What did you think would happen? (3 pts)
2. Explain your experimental design. In general, what was your procedure?
Charts, pictures, and diagrams are highly recommended. PLEASE do not list
every single step – that’s for your lab book. Simply give a “detailed overview”.
(3 pts)
3. Describe your results. Well-labeled pictures or charts are strongly
recommended. (3 pts)
4. Analyze your results. Think about your experimental question. What do you
conclude based on your results? What experiment would you suggest doing
next? (3 pts)
PRESENTATION NOTE #1: Your presentation should be somewhere between 5 –
7 slides. Please don’t plan on a presentation that will last longer than 10 minutes,
however, I don’t think you can do a good job on this in less than 5 minutes.
PRESENTATION NOTE #2: Your presentation should be saved to your student
folder and your partners’ student folder just in case they aren’t here when it is
time to give your presentation. YES, you will still present if your partners aren’t
here. You did the experiment, too, remember?
RESEARCH TIMELINE
Wednesday and Thursday 3/4 and 3/5 – Design experiment.
Friday 3/6 – Work on lab.
Monday 3/9 – Work on lab.
Tuesday 3/10 – Finish lab work (?) or meet in computer lab to design presentation.
Wednesday 3/11 – Presentations. Lab books due. Unit Test on Thursday 3/12.
GEL ELECTROPHORESIS/RESTRICTION ENZYME LAB GRADING RUBRIC
NAME:
EXPERIMENTAL QUESTION:
CATEGORY
NOTES
PTS
EARNED
PTS
POSSIBLE
Experimental Design – How well
does your experiment test your
experimental question?
4 pts
Scientific Work – How well do
4 pts
you run your experiment?
Lab book – Title, Experimental
question, Explanation of
Experimental question, Results,
Analysis
5 pts
Presentation – Experimental
Question: What was it? Scientific
importance? What did you think
would happen? Use a diagram to
explain your anticipated results.
3 pts
Presentation – Experimental
Design: What was your procedure?
Charts, pictures, diagrams. “Detailed
overview”
3 pts
Presentation – Results: Well-
3 pts
labeled pictures or charts.
Presentation – Analyze Results:
What do you conclude? Explain what
worked and what didn’t work. What
experiment would you suggest doing
next?
3 pts
TOTAL _______
/ 25 pts