Microsatellite marker based parentage evaluation in Murrah buffalo

Abstract

Accurate pedigree recording by DNA based method prevents any confusion and gives additional advantage to the conventional data recording process. For Bos tauras there are some molecular markers known for this purpose with reasonable accuracy. However, there is no such established markers for Bos bubalus bubalis to validate the recorded pedigree data. In this present study Murrah buffalo were considered for parentage evaluation. Thus a set of markers were developed to support the conventional pedigree record with objectives of optimization of number of microsatellite markers and development of microsatellite markers based methodology for parentage determination. Among 12 recommended markers by ISAG, four markers, ETH10, BM2113, ETH225, and TGLA227 were optimized for present study based on their reported PIC, readability, multiplexing, heterozygosity, and number of alleles by earlier workers. The markers were validated on three Murrah half sib families of 3-5 progenies from respective dams with common sire (LRC & ABRC, NDRI, Karnal). These markers were analysed in the genomic DNA isolated from blood and successfully amplified in selected families. Agarose gel electrophoresis (MetaPhor®, LONZA) successfully resolved alleles of all four optimized microsatellite markers in Murrah families. The results revealed locus BM 2113, ETH 225 and ETH 10 monomorphic in Murrah animals.