SNP (rs#) Primers Amplicon (bp) Annealing temp. (°C)/[MgCl2] (mmol) Restriction enzyme/allele size
T-786C (rs 2070744) promoter P1: 5’-GCA TGC ACT CTG GCC TGA AGT G-3’
P2: 5’-CAG GAA GCT GCC TTC CAG TGC-3’
223 61.5/2 MspI (Bioron)/T=162, 61 bp
C=116, 61, 46 bp
27 VNTR Intron 4   P1: 5’-AGG CCC TAT GGT AGT GCC TTT-3’
P2: 5’-TCT CTT AGT GCT GTG GTC AC-3’
393/420 59.2/3.5 a allele=393 bp
b allele=420 bp
G894T (rs 1799983) Exon 7   P1: 5’-AAG GCA GGA GAC AGT GGA TGG A-3’
P2: 5’-CCC AGT CAA TCC CTT TGG TGC TCA-3’
457 62.9/3 BanII (New England Biolabs Inc.)
G=137, 320 bp
T=457 bp
Table 1: Standard PCR conditions used in genotyping T-786C, G894T and 27VNTR SNPs of the eNOS gene.