shelf-life of 1st strand cDNA?

sullivan at gwu.edu sullivan at gwu.edu
Fri Apr 17 00:48:39 EST 1998



I'm doing lots of RT-PCR lately, and getting frustrated by
a recurring problem.  My first strand cDNA works fine the first
time I use it in PCR, and maybe the second, but subsequently the
I lose the ability to amplify rare messages out of it.  My RNA
looks good on a gel, extracted by guanidium methods; I store
the cDNA in the RT buffer (i.e., LTI Superscript II reaction buffer)
at -20 degrees.  Does an RT reaction product have a 'shelf life'?
Or should I look elsewhere for the cause of this problem?

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