Table 2 Primers and reaction conditions for mitochondrial DNA fragment amplification
No. | Primer | Sequence (5′-3′) | HC_F | SI_C | Annealing temperature | Extension time | ||
|---|---|---|---|---|---|---|---|---|
Position (bp) | Identity (%) | Position (bp) | Identity (%) | |||||
I | HSM1F | TAACGGATTAAAGCACAGCACTGAA | 9879-9903 | 96.00 | 9877-9901 | 100.00 | 61 °C | 3 min 30 s |
HSM1R | CGCATAGAGCTTGAAGGGAATTTAA | 13,094-13,118 | 96.00 | 13,089-13,113 | 100.00 | |||
II | HSM2F | TCTTGTTTTCTTGTTTTTGTGAGTT | 2812-2836 | 68.00 | 12,874-12,898 | 100.00 | 54 °C | 3 min |
HSM2R | CTCGTGTATCAACATCCATTCC | 3541-3562 | 63.64 | 886–907 | 95.45 | |||
III | HSM3F | TGCCATGATTGCAATAGGAGT | 825–845 | 85.71 | 825–845 | 100.00 | 57 °C | 3 min 30 s |
HSM3R | ATCTACAAAGTGTCAGTATCAGGCA | 4251-4275 | 92.00 | 4250-4274 | 100.00 | |||
IV | HSM4F | CCACTTCTCAACCCATCACCACTTT | 4212-4236 | 92.00 | 4211-4235 | 96.00 | 53 °C | 3 min 30 s |
HSM4R | CTATTCCTTGGGGGCCTATTTCTTC | 8060-8084 | 80.00 | 8058-8082 | 100.00 | |||
V | HSM5F | TTTTATCTCCTCCCTTTTTATYTCT | 8008-8032 | 76.00 | 8006-8030 | 96.00 | 55 °C | 2 min |
HSM5R | CCTCWAAAGTAGTTAAGATTGGGAC | 9955-9979 | 76.00 | 9953-9977 | 96.00 | |||
VI | HcM1F | ATCCTGCCTTCCGTTATTTTA | 9879-9903 | 96.00 | 9877–9901 | 100.00 | 53 °C | 2 min |
HcM1R | CAACAGTGGTTTGGTCCTTCT | 13,094-13,118 | 96.00 | 13,089-13,113 | 100.00 | |||
VII | RandomF | CCGCAAGGGAAAGATGAAATAC | 14,289-14,310 | 100.00 | 14,285-14,306 | 100.00 | 52 °C | 4 min |
RandomR | GGGGTGTTATTCTTCTAAGTATTGA | 2600-2624 | 84.00 | 2598-2622 | 100.00 | |||
VIII | HcM2F | GAAGGACAAGAACTGGAGAC | 2096-2116 | 100.00 | 1698-1717 | 75.00 | 53 °C | 3 min |
HcM2R | CTTTGCGAGATAGATTTAGC | 4851–4870 | 100.00 | 10,672-10,691 | 33.33 | |||
IX | HcM3F | ACTTTTGTTTTTCAATAAATCCCTCCA | 7680-7706 | 100.00 | 8631-8657 | 70.37 | 55 °C | 2 min 30 s |
HcM3R | TTTCTTTCTAACCACCCTTTTCACC | 10,229-10,253 | 100.00 | 10,230-10,254 | 88.00 | |||
X | RDF | TATCATTTAGTAGACCAAAGCCCAT | 3559-3583 | 100.00 | 3557-3582 | 100.00 | 52 °C | 40 s |
RDR | CCTGTAGCGACAAAGAAGGTAGAAC | 4118-4142 | 80.00 | 4117-4141 | 100.00 | |||