I have a lab quiz about bacteria transformation lab. There are some questions about transformation but I don’t know…. Can someone help me?
1. A student places pBC (the small p before the name always means a plasmid. Do not confuse with symbol for promoter, which is always a capital P with subscript of name of promoter, like Plac or Pbad) into E. coli. The plasmid contains the antibiotic resistance gene for chloramphenicol. What should they do to ensure they get transformants?
2. A student adds LB instead of calcium chloride to their tube by mistake. What effect will this most likely have on the transformation?
3. A student fails to heat shock their cells? They get transformants, but 10 times less than expected. What does this tell you about the role of “heat shock” in the transformation of E. coli.
4. What is 100 ml in µl? Which micropipette would you use for this, the p100 or p1000?
5. A student discovers that in their pGLO transformation the cells only glow under UV light on the LB/amp/ara plate? Why is this?
6. A student decides to clone the GFP gene into a plasmid under the control of the lactose promoter. Once they transform the plasmid into the cells, what will they have to add to turn the GFP gene on, i.e. produce GFP.
7. Match the following terms (May have more than one answer)
a. bla gene ___ 1. bacterial nutrients
b. ori ___ 2. transcription
c. LB broth ____ 3. replication of DNA
4. resistance to ampicillin
5. produces a lawn of E. coli
1. Use General perpose plasmid vector system for that. Generall purpose vectors usually include a system for detecting the precense of a cloned DNA fragment, based on the loss of an easily scored phenotype.The most widely used system involves the lacZ gene encoding the alpha-peptide from N-terminus of E-coli B-galactosidase.