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. Author manuscript; available in PMC: 2009 Aug 1.
Published in final edited form as: Pharmacogenomics. 2008 Oct;9(10):1445–1458. doi: 10.2217/14622416.9.10.1445

Table 1.

PCR and sequencing primers for assessing four VKORC1 SNPs by pyrosequencing.

SNP ID Primers Primer sequence
rs17886199 Forward 5′CCCCACTCCTAGCAATCTTGGTG3′
Reverse Biotin-5′CTCACTTTGGGCAACAGAGCCAG3′
Sequencing 5′GTCTTTTAATTGGTTTAAG3′
rs9923231 Forward 5′TGTTGGCCAGGCTTGTCTTAAAC3′
Reverse Biotin-5′AGCCAGCAGGAGAGGGAAATATC3′
Sequencing 5′GCGTGAGCCACCGCA3′
rs7196161 Forward 5′ATGTGAGACAGTCCCACTCTGC3′
Reverse Biotin-5′TGTAATCCCAGCACTTTAGGAAGCCA3′
Sequencing 5′ACTCCTGACCTCATGATC3′
rs17878338 Forward 5′CCGACCTCAGGTGATCTGCC3′
Reverse Biotin-5′CAGAGGCAGCTGTGGGTAAGG3′
Sequencing 5′CTGCTTTGGCCTCCC3′

PCR conditions for rs17878338 and rs9923231 were an initial denaturation of 96°C for 5 min followed by 35 cycles of 94°C for 20 s, 56°C for 20 s, and 72°C for 15 s. PCR conditions for rs17886199 were similar except that 60°C was used as the annealing temperature. Amplification of rs7196161 required a two-step ‘touchdown’ PCR reaction. Following denaturation, the annealing temperature was decreased 1°C every two cycles from 63°C down to 57°C. The second phase consisted of 30 cycles with a final annealing temperature of 56°C.