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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.

From: Oligonucleotide indexing of DNA barcodes: identification of tuna and other scombrid species in food products

Primer name

Primer sequence 5'-3'

PCR primer cocktails

FRAGMENT AB

USBotA-CytbsF

GCCAGT GATTACAACGGGGCCTCTTTCTTCTT

USBotB-CytbsF

GCCAGT GATTACAACGGCGCYTCCTTCTTCTT

USBotCbis-CytbsF

GCCAGT GATTACAACGGTGCTTCYTTCTTCTT

USBotDbis-CytbsF

GCCAGT GATTACAACGGGGCCTCCTTCTTCTT

USS1-UniT-R

TGACC CGATCGTRGCATTGTCTACTGARAAGCC

USS1-UniK-R

TGACC CGATCGTGGCGTTGTCTACTGAAAAGCC

USS1-UniR-R

TGACC CGATCGTGGCGTTGTCTACTGAGAAGCC

USS1-UniSA-R

TGACC CGATCGTRGCATTGTCTACAGAGAAGCC

Sequencing primers

 

USbot2-F

GTTGTAAAACGACGGCCAGT GATTACAACGG

USS2-UniT-R

CACAGGAAACAGCTATGACC CGATCGTRGC

PCR primer cocktails

FRAGMENT A

USBotA-CytbsF

GCCAGT GATTACAACGGGGCCTCTTTCTTCTT

USBotB-CytbsF

GCCAGT GATTACAACGGCGCYTCCTTCTTCTT

USBotCbis-CytbsF

GCCAGT GATTACAACGGTGCTTCYTTCTTCTT

USBotDbis-CytbsF

GCCAGT GATTACAACGGGGCCTCCTTCTTCTT

USAP-UniT-R

ATGACC GACCAAGGGAAGRACGTAGCCRACGAA

USAP-UniK-R

ATGACC GACCAAGGGAAGTACGTARCCGACGAA

USAP-UniR-R

ATGACC GACCAATGGRAGTACGTAGCCGACGAA

USAP-UniE-R

ATGACC GACCAAAGGGAGTACGTAACCGACGAA

USAP-UniSA-R

ATGACC GACCAAGGGAAGRACGTARCCGACGAA

Sequencing primers

 

USbot2-F

GTTGTAAAACGACGGCCAGT GATTACAACGG

USAP2-R

CACAGGAAACAGCTATGACC GACCAANGG

PCR primer cocktails

FRAGMENT B

USS1-UniT-F

GCCAGT GATTACTTCGTYGGCTACGTYCTTCCCT

USS1-UniK-F

GCCAGT GATTACTTCGTCGGYTACGTACTTCCCT

USS1-UniR-F

GCCAGT GATTACTTCGTCGGCTACGTACTYCCAT

USS1-UniE-F

GCCAGT GATTACTTCGTCGGTTACGTACTCCCTT

USS1-UniSA-F

GCCAGT GATTACTTCGTCGGYTACGTYCTTCCCT

USS1-UniT-R

TGACC CGATCGTRGCATTGTCTACTGARAAGCC

USS1-UniK-R

TGACC CGATCGTGGCGTTGTCTACTGAAAAGCC

USS1-UniR-R

TGACC CGATCGTGGCGTTGTCTACTGAGAAGCC

USS1-UniSA-R

TGACC CGATCGTRGCATTGTCTACAGAGAAGCC

Sequencing primers

 

USS2-UniT-F

GTTGTAAAACGACGGCCAGT GATTACTTCGT

USS2-UniT-R

CACAGGAAACAGCTATGACC CGATCGTRGC

  1. 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).
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BMC Biotechnology

ISSN: 1472-6750

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