As the semester ends, some students need extrinsic motivation. There is a 20 point Lab Performance score. Just do your work and score 20 points; or lose 20 points for not doing your work.
As we start to investigate this topic, there is some background information to explore.
Please complete this pre-lab as part of the lab Inquiry:
Use the web to read about the topics and to answer the questions below. There may be a quiz on this information and the lab procedure.
PRE-LAB QUESTIONS:
1. What is a use for "cleavage of DNA"?
2. We are using Tris-acetate-EDTA with a pH of 7.8 as our buffer. Describe its properties. Also, explain what is meant by pH, and a pH of 7.8
3. Define : Restriction Enzyme
4. What is meant by the 5 prime and 3 prime end of DNA? Find a diagram which shows this structure in DNA.
5. What is Bam H1? What organism is its source? What is it used to do?
6. What is the difference between a circular plasmid DNA and a linear DNA?
7. The lab is set up with Reaction buffer, DNA samples, water , and restriction enzymes in reaction tubes. What happens to the samples after the enzymes are added? What stops the reaction?
KNOW THE LAB SCHEDULE : This is a four day long lab. READ the lab procedure.
Day 1 : Micopipetting : ()
LABEL Reaction Tubes ( Name and Numbers) Load the samples into a reaction tube. Add DNA, D.I .water, and Buffer. DO NOT add the enzyme.
Day 2: The Reaction : ( Genetic engineering)
Add restriction enzymes to the reaction tubes. Incubate in a water bath -30 minutes. Add 5 uL 10x gel loading solution to each reaction tube. Prepare the Gels and Buffers for the chambers, if not yet completed.
Day 3 : Gel Electrophoresis Analysis ( This may be skipped, since it does not effect the outcome.)
1. Heat the samples in a water bath at 65 C.
2. Use micropipets to load the gels for electrophoresis, and run the gel.
Day 4 : Analyze the Results : ( Smell Assay - cell culture)
Stain the gels and Analyze the results.