Assignment 2

Plasmid isolation and plasmid features

Assignment must be submitted via turn-it-in

One of the goals of this project is to subclone GeneA from the pKC7 plasmid (pBR322 plasmid with GeneA inserted) into the pUC19 vector. In order to understand why you would want to do this, you need to understand the features of the two plasmids.

  1. How does the pUC18 vector differ from the pKC7 plasmid and how do these differences make it a better plasmid for subcloning?
  2. What feature(s) of pUC18 make it a better vector for production of GeneA?

The following questions are about the plasmid isolation protocol we used. Some answers can be found in the manufacture's manual. You may have to dig a little harder for others.

  1. In the GeneJET plasmid isolation protocol, where is the plasmid DNA located at the end of steps 4, 6, 7 and 10?
  2. If you are given a 1.5 ml tube of reagent and you have been told there is 10 ┬Ál in tube but you cannot see any liquid what might have happened and how can you fix the problem?
  3. What is the purpose of the GeneJET column?
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