Quickest method to visualize plasmid presence in bugs?
Philip L. Carl
plc at med.unc.edu
Thu Feb 1 09:47:11 EST 1996
In article <zGDRRIA8BgDxEwCv at genesys.demon.co.uk>,
"Dr. Duncan Clark" <duncan at genesys.demon.co.uk> wrote:
>In article <jpcd0-2901961926050001 at macr1-2.welc.cam.ac.uk>, John Dixon
><jpcd0 at mole.bio.cam.ac.uk> writes
>>Can anyone recommend a fast way to differentiate between bacterial
>>cultures/colonies that do or dont contain plasmids? Some sort of
>>boiling/alkaline lysis and immediate running perhaps.
We use the method of Sekar, Biotechniques 7:11-13 (1987). It takes a little
practice to know how much of the colony to sample, and the results are not
pretty because of all the bacterial DNA present in the lysate, but the
technique is fast and cheap. Post run staining with Syber Green makes the gel
look even better.
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