Hi Prabesh,

I understand that you have looked for literature on that and you have not found the primers. I think that this approach (http://www.biotechniques.com/multimedia/archive/00010/00295rr03_10055a.pdf, and following references) could suit you too. Other than that, since you are the first one to do that, why don't you just align the nucleotide sequences and design the primers yourself? I recommend primerBLAST to design specific primers.

Best,

Ruben

Thanks Ruben, we have decided to use Primer1 to design AS primers.... I hope it is reliable.

Hi, by chance I saw this article a few days ago: http://www.biotechniques.com/BiotechniquesJournal/2004/January/Detection-of-extremely-rare-alleles-by-bidirectional-pyrophosphorolysis-activated-polymerization-allele-specific-amplification-Bi-PAP-A-measurement-of-mutation-load-in-mammalian-tissues/biotechniques-116327.html

I guess that it is an overkill for what you want to do, but interesting anyway.

As to the primerBLAST (a NCBI service), now I see why you want to use the Primer1. Anyway, for other purposes, the big advantge of primerBLAST is that it designs the primers (using primer3), and then it blasts them against the reference genome to make sure that they are specific.

Cheers and good luck,

Ruben