Primer sequences and conditions used for PCR amplification
Primer Name | Sequence (5′-3′) | Conditions |
|---|---|---|
| Primers for MT1 | ||
| MT1-exon1-F1 | ACACGTGAATGAACAGCCTCGCTG | 95°C × 3 min, 1 cycle/95°C × 30 s, 61.8°C × 30 s, |
| MT1-exon1-R1 | GTCCCCATAGGAAAGAGAATGCGACTCT | 72°C × 30 s, 25 cycles/72°C × 10 min, 1 cycle |
| MT1-exon2-F1 | GCAGCTGTGACGTCAGGTCATCAG | 94°C × 2 min, 1 cycle/94°C × 15 s, 60°C × 20 s, |
| MT1-exon2-R1 | AAGCTGGAAGGTTCCTCCACCA | 68°C × 90 s, 20 cycles/68°C × 5 min, 1 cycle |
| MT1-exon1-F2 | AAGCGGGCTCGCGACGGA | 95°C × 3 min, 1 cycle/95°C × 30 s, 61.8°C × 30 s, |
| MT1-exon1-R2 | CGATGGGCAGGCTGGAGGAAGA | 72°C × 30 s, 25 cycles/72°C × 10 min, 1 cycle |
| MT1-exon2-F2 | GCAGGCAGAAGCAGCTCCTTCTT | 94°C × 2 min, 1 cycle/94°C × 15 s, 60°C × 20 s, |
| MT1-exon2-R2 | AGGCAACACGGAAAGGCGTG | 68°C × 90 s, 30 cycles/68°C × 5 min, 1 cycle |
| MT1-exon1-F3 | ATGCCGGGCAACGGCAGCGCGCTGC | 95°C × 3 min, 1 cycle/95°C × 30 s, 55°C × 30 s, |
| MT1-exon1-R3 | AAGATGTTTCCTGCGTTCCGGAGCTTCTTGTTCCG | 72°C × 30 s, 30 cycles/72°C × 10 min, 1 cycle |
| MT1-exon2-F3 | CGGAACGCAGGAAACATCTTTGTGGTGAGCTTAGC | 94°C × 2 min, 1 cycle/94°C × 15 s, 60°C × 20 s, |
| MT1-exon2-R3 | TTAAACCGAGTCCACCTTTACTAGA | 68°C × 1 min, 20 cycles / 68°C × 5 min, 1 cycle |
| MT1-exon1-F3 | 95°C × 3 min, 1 cycle / 95°C × 30 s, 55°C × 30 s, | |
| MT1-exon2-R3 | 72°C × 30 s, 30 cycles/72°C × 10 min, 1 cycle | |
| Primers for MT2 | ||
| MT2-F1 | CAGTACTGCGCGAGCCCTGCG | 94°C × 2 min, 1 cycle/94°C × 15 s, 68.4°C × 20 s, |
| MT2-R1 | CACCATTTCATGAGTTTGTCGTGC | 68°C × 90 s, 20 cycles/68°C × 5 min, 1 cycle |
| MT2-F2 | AAAGCGCAGCGCGGGAGAGT | 94°C × 2 min, 1 cycle/94°C × 15 s, 66.8°C × 20 s, |
| MT2-R2 | AGCTGGTGTGCCTCAGATCCAG | 68°C × 90 s, 30 cycles / 68°C × 5 min, 1 cycle |