PCR uusing 3 primers
 

Hello!!
I wanted to do a PCR using 3 primers in a mix. 1 is a redundant primer and the other are mating type specific primers. For a diploid wild type, i keep on getting only 1 band instead of the 2 expected band...plz help!!

Re: PCR uusing 3 primers
 

Maybe this "redundant" primer isn't annealing? Are the lengths of these primers similar or quite different? might want to try and find a "happy medium" annealing temp (I use 55 C) for all your primers so the they can all anneal properly.