BIMM100 Problem Set 2_JL_TAs

C a hospital is overloaded with newborns and four

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Unformatted text preview: by size via _____________________. C. A hospital is overloaded with newborns and four babies have been switched around to different incubators. Which baby (lanes 3-6) belongs to the parents (lanes 1 and 2)? 6. Gene Cloning A. What is the function of restriction enzymes? Are they endonucleases or exonucleases? B. What are the essential DNA sequences of a plasmid cloning vector? Explain the function of these elements. C. You decide to use the following plasmid to clone Gene A. i. You first digest both the genomic DNA and plasmid DNA using a restriction enzyme. Which restriction enzyme (NotI, HindIII, Sal1, or Bam HI) would you use to digest Gene A for insertion of the entire gene into the plasmid? Which restriction enzyme would you use to digest the plasmid before insertion of Gene A? Why? ii. You then plate these transformed bacterial cells onto media that allow you to distinguish between bacterial cells that obtained the plasmid and those that did not. Onto what type of growth medium will you plate your transformation mix? Why? 7. Gene Inactivation A. What are the general steps for gene inactivation in yeast? B. After many years of work, you have finally succeeded in creating a vector to make a conditional knockout mouse inserted into the germline. What were the general steps you took to get to this point? C. List two methods you can use to screen your mice for transmission of the vector with the loxP sites? Question 8. You wish to anneal the DNA primer to the single-stranded DNA template shown below. Single-stranded DNA template: 5’-TATAGCTACCATCGATAT-3’ DNA primer: 5’-CGATGGTA-3’ a) On the single-stranded DNA template, underline where the primer will anneal. (3) b) After annealing your primer, you add DNA polymerase and four nucleotides. Circle the types of nucleotides you would need to add in order to achieve DNA synthesis? (2) NMPs, NDPs, NTPs, dNMPs, dNDPs, or dNTPs c) Write, in the 5’-to-3’ direction, the product you would get after DNA synthesis from the primer (You do not need to write the DNA template, but inclu...
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