Primer sequences of CYP1B1 amplicons, PCR conditions, and DHPLC analysis parameters
| Amplicon | Primer sequences (5’ to 3’) | PCR product size (bp) | MgCl2 (mM) | Annealing temperature (°C) | Acetonitrile gradient (%B), start–end | Oven temperature (°C) |
|---|---|---|---|---|---|---|
| F, Forward, R, Reverse. | ||||||
| Exon 2_A | F: CGCTCCTGTCTCTGCACCCC | 302 | 0.5 | 60 | 57–63 | 65 |
| R: GGCGCGGCGCTACGGCGACG | 53–59 | 69 | ||||
| Exon 2_B | F: TGGCCACTGATCGGAAACGC | 456 | 1 | 57 | 61–67 | 67 |
| R: CCAACGTCATGAGTGCCGTG | 59–65 | 70 | ||||
| Exon 2_C | F: GCCACGTGCTGAGCGAGGCG | 329 | 1 | 57 | 58–64 | 65 |
| R: CATCCTGGACAAGTTCTTGA | 57–63 | 66.5 | ||||
| 52–58 | 71 | |||||
| Exon 2_D | F: GGTGCGCACCGTTTTCCGCG | 395 | 1 | 55 | 59–65 | 62 |
| R: GGGTTTGGGGTTTCGCTCCA | 57–63 | 64.5 | ||||
| Exon 3_B | F: GAGAACTTTGATCCAGCTCG | 343 | 1 | 55 | 58–64 | 58 |
| R: GCAAGAGGCAAGCTGAAATT | 55–61 | 60.5 | ||||