PCR dead ?

J.M. Ossewaarde JM.Ossewaarde at rivm.nl
Thu Apr 30 06:16:12 EST 1998


We also use Boehringer reagents. Last year and early this year we had
problems with their nucleotides. One batch showed very low RT-PCR
performance. Replacing this batch with a new batch solved our problems.

--
J.M. Ossewaarde, MD, PhD
Research Laboratory for Infectious Diseases
National Institute of Public Health and the Environment
PO Box 1
3720 BA Bilthoven
The Netherlands
Phone: +31 30 274 3942
Fax: +31 30 274 4449
Email: JM.Ossewaarde at rivm.nl

Thomas Kreuzer wrote in message <354776B5.6154 at kfunigraz.ac.at>...
>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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