From my experience this should work fine. I typically use 18 - 20 bp in my primers that bind to my gene of interest and in this case, a single mismatch should not really limit efficiency nor specificity. Anyway, to rule this out, run a gradient PCR and analyze your products on a gel. Then use the annealing temperature with the best result.
From my experience this should work fine. I typically use 18 - 20 bp in my primers that bind to my gene of interest and in this case, a single mismatch should not really limit efficiency nor specificity. Anyway, to rule this out, run a gradient PCR and analyze your products on a gel. Then use the annealing temperature with the best result.