PCR anomaly

CSC un691cs at genius.embnet.dkfz-heidelberg.de
Wed Oct 13 05:24:50 EST 1999


On Tue, 12 Oct 1999, Mark Edward Bowen wrote:

> My PCR reactions have suddenly become dominated by two well defined and
> reproduceable small MW (~500bp) products.  It happens with different
> buffers, enzymes, primers and templates.  The products always appear to
> be the same size.  I can anneal at temperatures up to 72 C and only lose
> my desired product.  I've tried hot start, touch down, Mg and
> temperature titrations as well as varying template and primer
> concentration.  I'm at a loss.  Any ideas?  I use a Stratagene
> robocycler.

Could you have it sequenced ? It sounds like fun.

Most likely you have a contamination. Clean the insides of your pipets
with DEPC water, throw out your batches of primers, exchange the water...

Kind regards,

Clemens Suter-Crazzolara, PhD

*************************************************************************
* For HUSAR, W2H & GCG visit:          http://genome.dkfz-heidelberg.de *
************************************************************************* 
* German             German Cancer Research Center DKFZ        European *
* EMBnet             Dept. Molecular Biophysics (H0200)            Data *
* node               INF 280, 69120 Heidelberg, Germany        Resource *
* - Genius -          phone ++49 6221-4223-42 fax -33           - EDR - *
*************************************************************************




More information about the Methods mailing list