In vivo labeling of PUC19 Help!!!
mleiblin at pop.ben2.ucla.edu
Sun Jun 22 11:03:03 EST 1997
Does anyone out there have a protocol for labeling PUC19 with
3H-Thymidine after electoporation into E. coli so that the
resultant labelled plasmid can be isolated in a suitable state for
subsequent electroporations. I would be interested in details such
as: 1) whether a minimal media is necessary to avoid the cold
thymidine in yeast based media; if so, which media and, 2) how much
labelled 3H-Thymidine in µCi should be used to produce an "adequate"
specific activity so that 25 ng of labelled PUC19 can be detected in
a ß counter.
Any assistance would be greatly appreciated
Thanks in advance
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