Genetic diversity between Murrah and Bhadawari breeds of Indian buffalo using RAPD-PCR
Abstract
The present study was conducted to estimate genetic diversity within and between Murrah and Bhadawari breeds using 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 series OPU and OPV, only 9 were found to be informative and were used further for amplification of genomic DNA. From the amplification profile of these primers, values of band frequency, genetic similarity, band sharing frequency, genetic distance, average percentage
difference and mean average percentage difference were calculated. From 9 random a total of 84 bands were amplified between breeds and out of these 51 were polymorphic (60.72%). In Murrah, overall percentage polymorphism of 56.36 was observed, while in Bhadawari it was 57.14. Average number of bands in Murrah was 5.04, while in Bhadawari were 5.03. Higher genetic similarity of 0.81 and 0.80 in Murrah and Bhadawari breeds, respectively, was observed as compared to between breed genetic similarity of 0.31. Genetic distance between breeds was 1.20. MAPD values of 10.72 and 16.10 were observed in Murrah and Bhadawari breeds, respectively, while MAPD of 60.73 was observed between breeds, indicating high genetic diversity between breeds. Six primers (OPU-01, OPU-02, OPU-05, OPU-07, OPU-14 and OPV-14) in Murrah and five primers (OPU-05, OPU-07, OPU-14, OPU-19 and OPV-14) in Bhadawari were found to be specific for these breeds.