Table 3 Details of the primers used for PCR amplification (A) Copy number and Relative expression studies by Real time PCR (B) in buffalo Bubalus bubalis
S.No. | Primer/Sequence ID | Forward | Reverse | Amplicon size (bp) | Annealing Temp (°C) | Nucleotide position of the primers in the sequence |
|---|---|---|---|---|---|---|
(A) PCR Primers | ||||||
1. | pDp1 | TGCACTCAATATGCCAGAAAA | TGAAAGCAGTCTGTTGTTCCA | 475 | 60 | 842-862 1296-1316 |
2. | pDp2 | TGCAGCAATCACAATCCTTG | AACATCCTGGTTGCTTCCAA | 124 | 60 | 66-85 170-189 |
3. | pDp3 | TTGCACAGAATGAGCAGTGGT | GGCATTAATACTGGGATCAGG | 101 | 60 | 492-512 572-592 |
4. | pDp4 | GGACAGAAATGGTATGGACCT | TATATTGGAGAAGGCGATGGC | 108 | 60 | 95-115 182-202 |
5. | BBACOT11 (a) | GAGCCGGCTGTCAGAGTC | AGCCAGACAACCTGGAATGT | 2504 | 60 | 34-51 2518-2537 |
6. | BBACOT11 (b) | TGAAGCTGAAGCCAACTGTG | TCCTAAAAAGATCTTCATGACCA | 2018 | 60 | 575-592 2570-2592 |
7. | BBACOT11g | AGTGAGGGGGTGTGCCCTGG | TGGTCGCACCCTCCTCGGAG | 601 | 60 | 1-20 582-601 |
8. | β-Actin | CAGATCATGTTCGAGACCTTCAA | GATGATCTTGATCTTCATTGTGCTG | 630 | 60 | 390-412 995-1019 |
9. | CD45 | GACATCGCAGTGTTTGTTGC | GGAGGTTCACATTCCTCTCG | 239 | 60 | 46-65 265-284 |
10 | CDH1 | TCTACAGCATCACTGGCCAACGAGCTG | TGCTTGGACCATCAGGGTGTATGTGGG | 476 | 60 | 566-592 1015-1041 |
(B) Real Time PCR Primers | ||||||
11. | Dp1 | CCTGATGTTCAAGCTGGTTTTAGA | CAGAGGATGTTGGCAATTTGATC | 65 | 60 | 1042-1065 1084-1106 |
12. | Dp2 | GCAGCAATCACAATCCTTGGA | TGTAGGTTTCTGTGTGGGCATAA | 63 | 60 | 67-87 107-129 |
13. | Dp3 | CAAAGCAGTGCACAAGAATCAAT | TGCCTTGCCCAACCTTTTTA | 63 | 60 | 318-340 361-380 |
14. | Dp4 | AAGACCAGGGATCTCTTCAAGAAA | CGAGCTCATCTTTGCATGAAAT | 66 | 60 | 24-47 68-89 |
15. | GAPDH | GCAAGTTCCACGGCACAGT | GATGGTGATGGCCTTTCCAT | 68 | 60 | 227-245 275-294 |