Primer sets for 2-step nested PCR and direct sequencing of mtDNA control region and CO1 and Cytb genes
Amplicon mtDNA region (np) (size, kb); mtDNA region (np) . | . | Sequence (5′ to 3′) for 2-step nested PCR . | . | Sequencing primers (5′ to 3′) . |
|---|---|---|---|---|
| 1 (1.12); control region (16024-16569; 1-576) | ||||
| F15574 (O) | CGCCTACACAATTCTCCGATC (O) | F15971 | TTAACTCCACCATTAGCACC | |
| R921 (O) | ACTTGGGTTAATCGTGTGACC (O) | R1 | CAGTGTATTGCTTTGAGGAGG | |
| F15971 (I) | TTAACTCCACCATTAGCACC (I) | SR | GCATGGAGAGCTCCCGTGAGTGG | |
| R1 (l) | CAGTGTATTGCTTTGAGGAGG (I) | SF | CATCTGGTTCCTACTTCAGGGTC | |
| 2 (1.39); Cytb (14688-15996) | ||||
| F 14622 (O) | CCACAAACCCCATTACTAAACCCAC (O) | F14688 | CTACAACCACGACCAATGATATG | |
| R16084 (O) | CGGTTGTTGATGGGTGAGTC (O) | R15996 | GCTTTGGGTGCTAATGGTGGAG | |
| F14663 (l) | GCATACATCATTATTCTCGCACGG (I) | |||
| R16057 (l) | GGGTGGTACCCAAATCTGCTTCC (I) | |||
| 3 (0.91); CO1 (6611-7470) | ||||
| F6541 (O) | GCAACCTCAACACCACCTTCTTCG (O) | F6645 | CTACCAGGCTTCGGAATAATCTCCC | |
| R7613 (O) | GTAGACCTACTTGCGCTGCATGTGC (O) | R7425 | GTTCTTCGAATGTGTGGTAGGGTG | |
| F6586 (l) | CCATTCTATACCAACACCTATTCTG (I) | |||
| R7498 (l) | CCATGGGGTTGGCTTGAAACCAGC (I) |
Amplicon mtDNA region (np) (size, kb); mtDNA region (np) . | . | Sequence (5′ to 3′) for 2-step nested PCR . | . | Sequencing primers (5′ to 3′) . |
|---|---|---|---|---|
| 1 (1.12); control region (16024-16569; 1-576) | ||||
| F15574 (O) | CGCCTACACAATTCTCCGATC (O) | F15971 | TTAACTCCACCATTAGCACC | |
| R921 (O) | ACTTGGGTTAATCGTGTGACC (O) | R1 | CAGTGTATTGCTTTGAGGAGG | |
| F15971 (I) | TTAACTCCACCATTAGCACC (I) | SR | GCATGGAGAGCTCCCGTGAGTGG | |
| R1 (l) | CAGTGTATTGCTTTGAGGAGG (I) | SF | CATCTGGTTCCTACTTCAGGGTC | |
| 2 (1.39); Cytb (14688-15996) | ||||
| F 14622 (O) | CCACAAACCCCATTACTAAACCCAC (O) | F14688 | CTACAACCACGACCAATGATATG | |
| R16084 (O) | CGGTTGTTGATGGGTGAGTC (O) | R15996 | GCTTTGGGTGCTAATGGTGGAG | |
| F14663 (l) | GCATACATCATTATTCTCGCACGG (I) | |||
| R16057 (l) | GGGTGGTACCCAAATCTGCTTCC (I) | |||
| 3 (0.91); CO1 (6611-7470) | ||||
| F6541 (O) | GCAACCTCAACACCACCTTCTTCG (O) | F6645 | CTACCAGGCTTCGGAATAATCTCCC | |
| R7613 (O) | GTAGACCTACTTGCGCTGCATGTGC (O) | R7425 | GTTCTTCGAATGTGTGGTAGGGTG | |
| F6586 (l) | CCATTCTATACCAACACCTATTCTG (I) | |||
| R7498 (l) | CCATGGGGTTGGCTTGAAACCAGC (I) |
CO1 indicates cytochrome c oxidase 1; Cytb, cytochrome b; np, nucleotide position; F, forward primer; O, outer primer; R, reverse primer; l, inner primer; SR, reverse sequence primer; SF, forward sequence primer.