multiple primers in PCR
cvasseur at icgm.cochin.inserm.fr
Fri Nov 28 06:40:34 EST 1997
In article (Dans l'article) <qtang-2611971253360001 at qtang.utmem.edu>,
qtang at nb.utmem.edu (sanglier) wrote (écrivait) :
> I'd like to use three different primer pairs to genotype my heterozygous
> double-mutant mice. The oligos are all roughly 20mers with similar Tm, and
> don't look like they will form dimers from a cursory check. The products
> are between 200 and 500bp. My question is, what concentration of each
> primer should I use? I had tried to use 1/3 of each to keep the final
> primer concentration the same as with single primers but had cruddy
> results. Is there anything else I should look out for?
> I'll be grateful for any suggestions!
You could have a look at :
" Multiplex PCR : Critical parameters and step-by-step protocol "
BioTechniques 23 : 504-511 (September 1997)
Hope this help!
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