multiple primers in PCR

sanglier qtang at nb.utmem.edu
Wed Nov 26 13:53:35 EST 1997


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!
Q



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