In vivo labeling of PUC19 Help!!!

Michael Liebling 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

Mickey Liebling



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