Primers used for MLST of Campylobacter jejuni/coli

Genes

The Campylobacter jejuni/coli MLST scheme uses internal fragments of the following seven house-keeping genes:

aspA (aspartase)

atpA (ATP synthase alpha subunit)

glnA (glutamine synthetase)

gltA (citrate synthase)

glyA (serine hydroxy methyl transferase)

pgm (phospho glucomutase)

tkt (transketolase)

PCR Amplification and sequencing

The primer pairs used for the PCR amplification and sequencing are detailed below:

aspA

Forward

aspAF1: GAGAGAAAAGCWGAAGAATTTAAAGAT

Reverse

aspAR1: TTTTTTCATTWGCRSTAATACCATC

atpA

Forward

atpAF: GWCAAGGDGTTATYTGTATWTATGTTGC

Reverse

atpAR: TTTAADAVYTCAACCATTCTTTGTCC

glnA

Forward

glnAF: TGATAGGMACTTGGCAYCATATYAC

Reverse

glnAR: ARRCTCATATGMACATGCATACCA

gltA

Forward

gltAF: GARTGGCTTGCKGAAAAYAARCTTT

Reverse

gltAR: TATAAACCCTATGYCCAAAGCCCAT

glyA

Forward

glyAF: ATTCAGGTTCTCAAGCTAATCAAGG

Reverse

glyAR: GCTAAATCYGCATCTTTKCCRCTAAA

pgm

Forward

pgmF1: CATTGCGTGTDGTTTTAGATGTVGC

Reverse

pgmR1: AATTTTCHGTBCCAGAATAGCGAAA

tkt

Forward

tktFN: AACGGTTTTAAGTTATCAT

Reverse

tktRN: TTTTGTGACTTCCTTCAAGC

Reaction conditions - PCR

Denaturation: 94ºC 30 s

Annealing: 53ºC 30 s

Extension: 72ºC 2 minutes

30 cycles