GeneReleaser and PCR on Whole Blood

Gary K. gtk10583 at glaxo.com
Wed Aug 23 11:43:31 EST 1995


This question is for anyone using BioVenture's GeneReleaser for PCR on
mouse whole blood.  I typically place one drop (approx. 5 ul) mouse whole
blood into an eppie containing 15 ul heparin solution (16 U/ml in sterile
water).  Then I use 1 ul for PCR analysis.  For the first week the assays
went very well with sharp specific bands on a gel.  However, the following
weeks lead to progressively more smearing of the PCRs until the diagnostic
band was no longer visible.  Could this be due to the heparin solution
going bad?  Should the heparin be stored at 4C instead of room temp as
indicated on the stock bottle (exp. 4/97)?  And would it be better to
prepare the working solution (16 U/ml) in PBS instead of water?

Thanks for any responses.
Gary



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