Table 2.
Primer cocktail (F, forward and R, reverse) and sequencing primers to detect polymorphisms occurring in fragments AB (226 bp), A (109 bp) and B (95 bp) of mitochondrial cytochrome b in 17 fish species of the Scombridae fish family.
| Primer name | Primer sequence 5'-3' |
|---|---|
| PCR primer cocktails | FRAGMENT AB |
| USBotA-CytbsF | GCCAGTGATTACAACGGGGCCTCTTTCTTCTT |
| USBotB-CytbsF | GCCAGTGATTACAACGGCGCYTCCTTCTTCTT |
| USBotCbis-CytbsF | GCCAGTGATTACAACGGTGCTTCYTTCTTCTT |
| USBotDbis-CytbsF | GCCAGTGATTACAACGGGGCCTCCTTCTTCTT |
| USS1-UniT-R | TGACCCGATCGTRGCATTGTCTACTGARAAGCC |
| USS1-UniK-R | TGACCCGATCGTGGCGTTGTCTACTGAAAAGCC |
| USS1-UniR-R | TGACCCGATCGTGGCGTTGTCTACTGAGAAGCC |
| USS1-UniSA-R | TGACCCGATCGTRGCATTGTCTACAGAGAAGCC |
| Sequencing primers | |
| USbot2-F | GTTGTAAAACGACGGCCAGTGATTACAACGG |
| USS2-UniT-R | CACAGGAAACAGCTATGACCCGATCGTRGC |
| PCR primer cocktails | FRAGMENT A |
| USBotA-CytbsF | GCCAGTGATTACAACGGGGCCTCTTTCTTCTT |
| USBotB-CytbsF | GCCAGTGATTACAACGGCGCYTCCTTCTTCTT |
| USBotCbis-CytbsF | GCCAGTGATTACAACGGTGCTTCYTTCTTCTT |
| USBotDbis-CytbsF | GCCAGTGATTACAACGGGGCCTCCTTCTTCTT |
| USAP-UniT-R | ATGACCGACCAAGGGAAGRACGTAGCCRACGAA |
| USAP-UniK-R | ATGACCGACCAAGGGAAGTACGTARCCGACGAA |
| USAP-UniR-R | ATGACCGACCAATGGRAGTACGTAGCCGACGAA |
| USAP-UniE-R | ATGACCGACCAAAGGGAGTACGTAACCGACGAA |
| USAP-UniSA-R | ATGACCGACCAAGGGAAGRACGTARCCGACGAA |
| Sequencing primers | |
| USbot2-F | GTTGTAAAACGACGGCCAGTGATTACAACGG |
| USAP2-R | CACAGGAAACAGCTATGACCGACCAANGG |
| PCR primer cocktails | FRAGMENT B |
| USS1-UniT-F | GCCAGTGATTACTTCGTYGGCTACGTYCTTCCCT |
| USS1-UniK-F | GCCAGTGATTACTTCGTCGGYTACGTACTTCCCT |
| USS1-UniR-F | GCCAGTGATTACTTCGTCGGCTACGTACTYCCAT |
| USS1-UniE-F | GCCAGTGATTACTTCGTCGGTTACGTACTCCCTT |
| USS1-UniSA-F | GCCAGTGATTACTTCGTCGGYTACGTYCTTCCCT |
| USS1-UniT-R | TGACCCGATCGTRGCATTGTCTACTGARAAGCC |
| USS1-UniK-R | TGACCCGATCGTGGCGTTGTCTACTGAAAAGCC |
| USS1-UniR-R | TGACCCGATCGTGGCGTTGTCTACTGAGAAGCC |
| USS1-UniSA-R | TGACCCGATCGTRGCATTGTCTACAGAGAAGCC |
| Sequencing primers | |
| USS2-UniT-F | GTTGTAAAACGACGGCCAGTGATTACTTCGT |
| USS2-UniT-R | CACAGGAAACAGCTATGACCCGATCGTRGC |
The amplicon size is respectively of 291 bp (fragment AB), 175 bp (fragment A) and 163 bp (fragment B). Italic: 5' Partial Universal Primer (PUP) and 5' Universal Primer (UP); bold: Read Start signal (RS1 and RS2); normal text: 3' cocktail portion (CP).