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.