Genotyping assay conditions for CYP3A5 and CYP3A7 allelic variants
Lowercase letters represent mismatches incorporated into primers to introduce restriction sites, indicated by bold letters.
Allele | Primers (5′ to 3′) | Annealing Temperature | Restriction Enzyme | RFLP Weight Pattern | RFLP Variant Pattern | ||
|---|---|---|---|---|---|---|---|
| °C | bp | ||||||
| 3A5*2 | F | CCT ACC TAT GAT GCC GTG G | 56 | DdeI | 105+66+33 | 138+66 | |
| R | AGT ACT TTG GGT CAT GGT GAA GAG CcT AA | ||||||
| 3A5*3 | F | CAT GAC TTA GTA GAC AGA TGA C | 55 | SspI | 148+125+20 | 168+125 | |
| R | GGT CCA AAC AGG GAA GAa ATA | ||||||
| 3A5*4 | F | TCG ACT CTC TCA ACA ATC CtC | 55 | TaqI | 261+20 | 241+20+20 | |
| R | AAA GTG TGT GAG GGC TCT cGA | ||||||
| 3A5*5 | F | CCA TGA AGA TCA CCA CAA CT | 53 | NlaIII | 226+14 | 189+37+14 | |
| R | CCT GTC CCC AGA TTC ATg C | ||||||
| 3A5*6 | F | CAA CAA TCC ACA AGA CCC CTT TGT GGA GAG Cct TAA | 58 | AflII | 273+31 | 304 | |
| R | GGC AGT GGG GTT TGT GGT GGG GTG | ||||||
| 3A5*7 | F | CTT GAA TAT CTC CTA CAT TCt TAA gTC C | 60 | AflII | 236+20 | 207+20+29 | |
| R | CTC CAT CTG TAC CAC GGC ATC ATA ctT A | ||||||
| 3A5 | F | GAT TTA CCT GCC TTC AAT TTT TCA CTG ACC | 70 | ||||
| 8152-bp template | R | GGC AGT GGG GTT TGT GGT GGG GTG | |||||
| 3A7*1C | F | ATG ACC TAA GAA GAT GGA GTG | 55 | SspI | 1617+585+266 | 2202+266 | |
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| R | GAA GGG CAT GGT CTA CAC TAT |
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F, forward; R, reverse.