Download Amgen Biotech Labs Pre-lab questions

Amgen Biotech Labs Pre-lab questions
www.bwbiotechprogram.com Click on Curriculum, then Student Manual (This link is
also on my teacherweb page)
Lab 1 Pipetting practice
1. What is the purpose of this lab?
2. What is 1ug equal to?
3. Why do we use such small quantities?
4. How much does a pound of DNA cost and why is it so expensive?
5. What volume is a P-20 micropipettor calibrated to dispense?
6. Read and summarize the precautions on page 1.2-1.3.
7. Summarize the procedure steps #1-3.
8. Copy the table from page 1.4 into your lab book.
9. At what steps do you put on a fresh tip?
Lab 1 Using Gel Electrophoresis to Separate Molecules
1. What is gel electrophoresis designed to do?
2. What separates the molecules?
3. Why do they move?
4. What is agarose made of? What is the function of the sodium borate?
5. Which end of the gel box is positive? Which is negative?
6. How much of each sample do you load?
7. Why does the sample sink into the wells?
8. How long do you run your gel?
Lab 2 Restriction Analysis of pARA and pKAN-R (pg 2.1)
1. What are plasmids? Where are they found?
2. What has been engineered into our plasmids and why does this function as a
selectable marker?
3. What gene does the pARA plasmid contain?
4. How big is the pARA plasmid? What does the gene do and what does it allow?
5. What are the sites “BamH1” and HindIII”?
6. What is the rfp gene? How big is it?
7. What does cutting pKAN-R with BamH1 and HindIII do?
Lab 2a pARA-R Restriction Digest
1. What do restriction enzymes do?
2. With plasmids, what can restriction enzymes be used to create?
3. In what organisms were restriction enzymes discovered? What do they do to
viral DNA?
4. What is left at the end of the restriction fragments?
5. Why can fragments cut by the same restriction enzymes anneal together?
6. Copy the table.
7. What is the purpose of adding water to the A- tube?
8. When do you put a fresh tip onto the pipettor?
9. Why do you put the tubes into the centrifuge?
Lab 4a Confirmation of pARA-R Restriction Digest
1. What is the purpose of this lab?
2. Why does DNA migrate through the gel?
3. Why can more than 1 band appear in the undigested plasmid lane?
4. What 3 types of plasmids might you see? Describe them.
5. Why do we add the loading dye?
6. Why do you pump the solution?
7. Why must you use a new tip for each sample?
8. At which end of the gel box should the wells be?
9. What volume of DNA size marker do you add to the gel?
10. How do you know if you’ve loaded the sample correctly?
11. How much A+ and A- do you load?
12. What voltage do you set the power supply to?
13. How do you know your gel is running? How long do you run it?
Lab 5 Transformation of E. coli with pARA
1. Define transformation
2. How do most bacteria pass on extra genetic material?
3. What can plasmids do for bacteria?
4. What factors determine transformation efficiency?
5. What does it mean for a cell to be “competent”? How is it done in the lab?
6. How does adding calcium chloride make it easier for the plasmid to get into
the cell?
7. What does heat-shocking the bacteria do?
8. What is in each of the 3 plates?
9. What does ampicillin do to bacteria? What if the bacteria has the ampr
gene?
10. What is arabinose do and what is its function in this lab?
11. To which tube do you add the plasmid?
13. Which half of the petri dish do you mark?
14.How much of each bacteria sample do you put onto each half of the LB and
LB/amp plates?
15. Which bacteria do you put onto the LB/amp/ara plate? How much?