Allele Specific and tetra-primer ARMS PCR Assays for Rapid Screening of SNPs at Male-specific Region of Y-chromosome in Cattle

摘要

Markers on the male-specific region ofY- chromosome (MSY) in mammals, paternally inherited in a haploid way, have widely been used to study the origin of species, range expansion, admixture of populations and migration in animals. Molecular variationin the Y chromosome provides information about genetic diversity, since it reveals the distribution of paternal lineages. Recent discovery of five polymorphic sites on cattle Y-chromosome genes (DDX3Y, UTY and ZFY) ailcaus identifying three haplotypes (Yl, Y2 and Y3) in cattle. Here we report the optimization and validation of allele specific PCR and tetra-primer ARMS PCR protocol for screening three SNPs ongenes (DDX3Y -intron 1,425>C/T; ZFY-intron 9,120> C/T; and ZFY- intron 10, 655> C/T) located at MSY of bull Y-chromosome. Although, both the techniques are found to be efficient in exploring Y-SNPs variation; however we propose to utilize allele specific PCR over and tetra-primer ARMS PCRfor screening SNPs at MSY ofY- chromosome in cattle as it requires little standardization and thereby less time consuming. Tlie standardized protocols may be useful for large scale screening of those Y-SNPs and analysis ofY-haplotypes in diverse native cattle breeds, exotic and crossbred cattle populations with high accuracy and reliability.