Genetic diversity and characterization of buffalo breeds by RAPD-PCR
Abstract
The present study was conducted for breed characterization and genetic diversity within and between Murrah, Bhadawari and Nili-Ravi breeds by RAPD-PCR. Genomic DNA was isolated from 20 unrelated animals of each breed. DNA was evaluated for quality, purity and concentration. Optimization of PCR was carried out using various concentrations of different components of reaction mixture. Out of 40 random primers of operon series OPU and OPV, only ten were found informative and were used further for amplication of genomic DNA. From the amplification profile of these primers values of band frequency, genetic distance, genetic similarity, band sharing frequency, average percentage difference and mean average percentage difference were calculated from ten random primers a total of 188 bands were amplified between as in three breed and out of these 112 were polymorphic (59.57%). In Murrah,
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Bhadawari and Nili-Ravi overall percentage polymorphism of 65.51, 63.49 and 53.44 observed, respectively. Higher genetic similarity of 0.84, 0.77 and 0.70 in Murrah, Bhadawari and Nili-Ravi, respectively. MAPD value of 11.41, 13.62 and 18.53 were observed in Murrah, Bhadawari and Nili-Ravi while MAPD of 69.0, 67.0 and 40.0 were observed between Bhadawari and Nili-Ravi, Bhadawari and Murrah and Murrah and Nili-Ravi, respectively. These values indicated that higher genetic diversity between breeds. Five primers (OPU-05, OPU-14, OPU-19, OPV-14, OPV-20) resolved Murrah specific bands while five primers gave Bhadawari specific bands viz. (OPU-01, OPU-05, OPU-07, OPU-14 and OPV-14) and five primers gave Nili-Ravi specific bands viz. (OPU-02, OPU-05, OPU-19, OPV-01, and OPV-14) were found to be specific for these breeds.