Journal
ELECTROPHORESIS
Volume 29, Issue 11, Pages 2419-2423Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/elps.200700702
Keywords
allele-specific primers; biallelic marker; mismatch; single nucleotide polymorphisms
Ask authors/readers for more resources
We describe the application of allele-specific PCR (AS-PCR) for screening biallelic markers, including SNPs, within the nonrecombining region of the human Y-chromosome (NRY). The AS-PCR method is based on the concept that the perfectly annealed primer-template complex is more stable, and therefore, more efficiently amplified under the appropriate annealing temperature than the complex with a mismatched 3'-residue. Furthermore, a mismatched nucleotide at the primer's 3'-OH end provides for a poor extension substrate for Taq DNA polymerase, allowing for discrimination between the two alleles. This method has the dual advantage of amplification and detection of alleles in a single expeditious and inexpensive procedure. The amplification conditions of over 50 binary markers, mostly SNPs, that define the major Y-haplogroups as well as their derived lineages were optimized and are provided for the first time. In addition, artificial restriction sites were designed for those markers that are not selectively amplified by ASTCR. Our results are consistent with allele designations derived from other techniques such as RFLP and direct sequencing of PCR products.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available