Table 2.

Primer sequences, thermocycling conditions, restriction enzymes, and gel variables used to genotype variants in CYP1A1, CYP1A2, CYP1B1, COMT, GSTM1, and GSTT1

PolymorphismPrimer sequencesThermocycling conditionsRestriction enzyme (5 units); gel variablesReferences
CYP1A1 m15′-GGCTGAGCAATCTGACCCTA-3′, 5′-GGCCCCAACTACTCAGAGGCT-3′95°C, 2 min; 35 × 94°C, 1 min; 58°C, 30 s; 72°C, 1.5 minMspI, SphI; 4% NuSieve GTG (FMC, Rockland, ME)(59)
(Triplex PCR) CYP1A1 m2 or m45′-GAAAGGCTGGGTCCACCCTCT-3′, 5′-CCAGGAAGAGAAAGACCTCCCAGCGGGCCA-3′72°C, 5 min; 95°C, 2 min; 35 × 94°C, 1 min; 60°C, 1 min; 72°C, 2 min; 72°C, 5 minNcoI, HinfI; 2.5% agarose (Invitrogen, Carlsbad, CA)(59)
GSTM1 (deletion)5′-CTGCCCTACTTGATTGATGGG-3′, 5′-CTGGATTGTAGCAGATCATGC-3′
GSTT1 (deletion)5′-TTCCTTACTGGTCCTCACATCTC-3′, 5′-TCACCGGATCATGGCCAGCA-3′
CYP1A1 m25′-GAAAGGCTGGGTCCACCCTCT-3′, 5′-CCAGGAAGAGAAAGACCTCCCAGCGGTC-3′95°C, 2 min; 35 × 94°C, 1 min; 60°C, 1 min; 72°C, 2 min; 72°C, 5 minHincII; 4% NuSieve GTG (FMC, Rockland, ME)This study
CYP1A1 m45′-GAAAGGCTGGGTCCACCCTCT-3′, 5′-GGCCCCAACTACTCAGAGGCT-3′95°C, 2 min; 35 × 94°C, 1 min; 62°C, 30 s; 72°C, 1.5 min; 72°C, 5 minBsaI; 2.5% agarose (Invitrogen, Carlsbad, CA)(59)
CYP1A25′-CAACCCTGCCAATCTCAAGCAC-3′, 5′-AGAAGCTCTGTGGCCGAGAAGG-3′95°C, 2 min; 40 × 94°C, 1 min; 62°C, 30 s; 72°C, 1 min; 72°C, 5 minApaI; 2.5% agarose (Invitrogen, Carlsbad, CA)(40)
CYP1B15′-TAAGAATTTTGCTCACTTGC-3′, 5′-GTTCTCCGGGTTAGGCCACTTAA-3′95°C, 2 min; 15 × 94°C, 1 min; 60°C, 30 s; 72°C, 1 min; 25 × 94°C, 1 min; 50°C, 30 s; 72°C, 1 min; 72°C, 5 minAflII; 4% NuSieve GTG (FMC, Rockland, ME)(76)
COMT Val158Met5′-TACTGTGGCTACTCAGCTGTGC-3′, 5′-GTGAACGTGGTGTGAACACC-3′95°C, 2 min; 40 × 94°C, 1 min; 62°C, 1 min; 72°C, 2 min; 72°C, 5 minNlaIII; 4% NuSieve GTG (FMC, Rockland, ME)(77)