Use this to make a map of restriction sites on your plasmid with insert for t

Apr 12th, 2015
Anonymous
Category:
Biology
Price: $20 USD

Question description

Please see attachment to complete questions:

Analysis Questions:

1.  Compare your DNA sample A with DNA sample B.  How are they the same? How are they different?

2.  How do you account for the differences? 

3.  The addition of neutralization buffer in during the isolation of the plasmid DNA causes the bacterial chromosomal DNA to precipitate with the white, soapy mixture that you spin into the bottom of the tube.  The plasmid DNA remains in the aqueous top layer when this white mixture is spun down.  What might be the consequence of using too MUCH bacteria?

4.  What happens when the lysis buffer is added to the bacterial solution?

5.  The plasmid-containing solution is loaded into the column, then washed, and then the plasmid is eluted with sterile water.  What is the importance of the resin that is added to the plasmid mixture?  How does the resin work?

6.  The DNA is visible under ultraviolet light after gel electrophoresis because the gel contains a fluorescent salt called ethidium bromide (which is a powerful mutagen and should NOT be allowed to touch your skin).  This salt wedges (intercalates) between the two helices along the length of the DNA strand.  Based on this explanation, which pieces of DNA should be most visible on the gel?  Which should be least visible?

7.  If a sample contained only bacterial DNA which had been sheared randomly into different sized pieces, how would it appear on the gel after electrophoresis and staining with ethidium bromide?

plamidquest.pdf 

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