RAPD-PCR based characterization of Murrah and Nili Ravi breeds of buffalo

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Date
2005
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LUVAS
Abstract
The present study was conducted to estimate genetic relatedness within and between Murrah and Nili Ravi breeds using RAPD-PCR. Genomic DNA was isolated from 20 animals of each breed, evaluated for purity, concentration and quality. Optimisation of PCR was carried out by varying the concentration of different components of the reaction mixture. A total of 11 decamer primers were employed for estimating band frequency, genetic identity index, band sharing frequency, genetic distance and mean average percentage difference between and within breeds. All the 11 primers amplified the genomic DNA, generating bands ranging from 150- 3000 bp. From the 11 random primers, a total of 110 bands were amplified and 78 of these (about 71%) were found to be polymorphic in Murrah breed. The number of polymorphic loci ranged from 2 to 13 in Murrah breed with an average of 7.10. From the 8 polymorphic random primers, a total of 57 bands were amplified and 34 of these (about 60%) were found to be polymorphic in Nili Ravi breed. The number of polymorphic loci ranged from 1 to 10 in -xxxvi- Nili Ravi breed with an average of 4.86. A total of 114 bands were amplified from the 11 random primers and 56 of these (about 49%) were found to be polymorphic between Murrah and Nili Ravi breeds. The number of polymorphic loci ranged from 2 to 11 between the breeds with an average of 5.82. Higher genetic similarity of 0.79 and 0.85 in Murrah and Nili Ravi breeds, respectively, was obtained as compared to between breeds genetic similarity of 0.62±0.06. Low overall genetic distance of 0.26 and 0.18 was observed in Murrah and Nili Ravi breeds, respectively, in comparison to between breeds genetic distance of 0.53±0.11. The MAPD in Murrah breed was 21.29% and in Nili Ravi breed was 14.57%. Between breeds MAPD was 36.64±5.85% indicating medium level of genetic diversity. Four random primers showed Murrah breed specific amplicons.
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