multiple primers in PCR
Dr Andrew Doherty
a.doherty at bris.ac.uk
Thu Nov 27 06:49:42 EST 1997
sanglier wrote in message ...
>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!
I just used the same concentration of primers as if there was only one
pair - 1uM each primer. I was doing two pairs - I would imagine the same
goes for three
Hope it helps
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