DNA amplification fingerprinting of cattle and buffalo genome by RAPD-PCR utilizing arbitrary oligonucleotide primers

Ganai, T.A.S.; Singh, R.K.; Butchiah, G.

Buffalo Journal 16(3): 331-339

2000


Accession: 003397770

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Abstract
For amplification of genomic DNA of cattle (Bos indicus) and buffalo (Bubalus bubalis) with short arbitrary oligonucleotide primers, four primers with the sequence, 5' GTGACGTAGG-3' (G1), 5'-TGCCGAGCTG-3' (G2), 5'-GTGGTGGTGG-3 (G4) and 5'-GCGAGCGTCCC-3' (C7), were screened. The two primers, C7 and G4, didn't yield any amplification product in PCR with templates of cattle and buffalo genomic DNA even after changing reaction conditions indicating that probably the sequences of these two primers didn't anneal with target DNA. But remaining two primers, G1 and G2, yielded the amplification product in cattle as well as buffalo genomic DNA and exhibited highly polymorphic patterns. The average number of bands/animals observed were 6.00 +or- 0.82, 1.00, 6.40 +or- 0.89 in buffalo and 6.75 +or- 1.10, 2.33 +or- 0.33, 8.50 +or- 0.71 in cattle, with primers G1, G2 and G1 + G2, respectively. The band sharing and probability of two individuals to exhibit identical fingerprints in the buffalo, as compared to cattle, were considerably low, and the estimates were 0.572 +or- 0.106, 0.514 +or- 0.060; 8.7 x 10-4, 1.85 x 10-4 in buffalo with G1 and G1 + G2 and 0.723 +or- 0.125, 0.788 +or- 0.050, 0.820 +or- 0.036, and 4.3 x 10-3, 3.0 x 10-1 and 2.6 x 10-2 in cattle with primers G1, G2 and G1 + G2, respectively.