Table 2 Primers and conditions used in rs2234922 (A > G), rs1051740 (T > C) and rs1800566 (G > A) genotyping
Description | General Characteristics |
|---|---|
EPHX1 | rs2234922 (Missense) |
Type of polymorphism | Single-base A > G |
Site of polymorphism | His139 Arg |
PCR primers | |
Forward | 5`- AGG GTG GCA GGA CTC AAT ATC TA −3` |
Reverse | 5`- AGG ATG CCT CTG AGA AGC CAT AG −3` |
PCR conditions | |
Denaturation | 95 °C, 5 min |
Annealing | 58 °C, 20 s |
Extension | 72 °C, 15 min |
No. of cycles | 30 |
Restriction enzyme | Csp6I (CviQI), (G ↓ TAC sequence) |
Restriction Enzymes Product Size (bp) | AA (319 bp) |
AG (153, 166 and 319 bp) | |
GG (153 and 166 bp) | |
EPHX1 | rs1051740 (Missense) |
Type of polymorphism | Single-base T > C |
Site of polymorphism | Tyr113His |
PCR primers | |
Forward outer primer | 5`- GTT TTC TGG AAA CAG ACT TTG CTC TTG T −3` |
Reverse outer primer | 5`- GAT TTG TTG TGA CTG CCA CCA TAT TTT TGCA −3` |
Forward inner primer (T allele) | 5`- AGA AGC AGG TGG AGA TTC TCA ACA TAT −3` |
Reverse inner primer (C allele) | 5`- ACC TTC AAT CTT AGT CTT GAA GTG AGG TTG −3` |
PCR conditions | |
Denaturation | 95 °C, 5 min |
Annealing | 58 °C, 30 s |
Extension | 72 °C, 30 s |
No. of cycles | 35 |
Product Size (bp) | T (234 bp) |
C (261 bp) | |
Common (436 bp) | |
NQO1 | rs1800566 (Missense) |
Type of polymorphism | Single-base G > A |
Site of polymorphism | P187S |
PCR primers | |
Forward outer primer | 5`- ATT CTC TAG TGT GCC TGA GGC CTC CT −3` |
Reverse outer primer | 5`- TGC CTG GAA GTT TAG GTC AAA GAG GCTG −3` |
Forward inner primer (G allele) | 5`- GCA TTT CTG TGG CTT CCA AGT CTT AGC AC −3` |
Reverse inner primer (A allele) | 5`- GTG CCC AAT GCT ATA TGT CAG TTG CGA −3` |
PCR conditions | |
Denaturation | 95 °C, 5 min |
Annealing | 57 °C, 30 s |
Extension | 72 °C, 30 s |
No. of cycles | 35 |
Product Size (bp) | G (176 bp) |
A (196 bp) | |
Common (316 bp) |