I have used CAPS markers (chu et al., 2009) to differentiate homozygous and heterozygous peanut population.
I have used different protocol and condition but the desire band not amplified.
After that I checked DNA, PCR chemicals, CAPS Primers everything is ok.
But still i am not getting amplification.
have you checked the primer sequences directly against your template sequence?
Sometimes primer sequences are not accurately reported. I would run a gradient pcr over Tm-6c to about Tm +5 c and also check your sequence for polymorphisms near the 3' end of each primer in your species and population. If both of those are not helpful get the primers re synthesised and try again as they may have been dissolved in water and become depurinated over time
Hi, have you checked the amplification with a different primer? This just to confirm that the DNA is really working. Good luck
check the F primers and R primers and the purity of your DNA then if its good the problem with thermal cycle. Good luck
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