PCR dead ?

Thomas Kreuzer thomas.kreuzer at kfunigraz.ac.at
Wed Apr 29 13:51:33 EST 1998


I have been measuring c-fos gene expression with RT-PCR
for almost 3 years now and never had problems with the
amplification (except in case of mistakes of course :-))

I have mostly worked with reagents from Boehringer, but
I have also used polymerases from other companies. 
I have used primers from different sources - everything 
worked.

But when I tried a new ready-to-use PCR mix a few weeks
ago, I could not get any amplification for the fos-samples
although the reaction worked for a standard (other primers).
I tried new primers - nothing.
I switched back to self-mixed reagents - nothing.
In the end, I did not even get amplification from old PCR
products (which I used as positive controls), no matter which 
reagents or which batch of primers I used. All the time 
the amplification of the standard worked.

I have run out of explanations now.
But maybe one of you can give me a hint.

Thanks in advance,
Tom
-- 
-----------------------------------------------
Tom Kreuzer
Institute for Biochemistry,K-F-University Graz,
http://bkfug.kfunigraz.ac.at/~kreuzer



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