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?
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