Primers used for MLST of Campylobacter upsaliensis

Genes

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

adk (adenylate kinase)
aspA (aspartase)
atpA (ATP synthase alpha subunit)
glnA (glutamine synthetase)
glyA (serine hydroxy methyl transferase)
pgi (glucose-6-phosphate isomerase)
tkt (transketolase)

PCR Amplification and sequencing

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

adk
Forward
adkF: TGAAAGAATTRTTTTTAATCATAGG
Reverse
adkR: CTTTCATRTCWGCHACGATAGGTTC

aspA
Forward
aspAF2: GAAGCWAAAGCWAAAGAATAYAAAGAT
Reverse
aspAR2: GAGTTTTTTGCAWGCTTCWGGATT

atpA
Forward
atpAF: GWCAAGGDGTTATYTGTATWTATGTTGC
Reverse
atpAR: TTTAADAVYTCAACCATTCTTTGTCC

glnA
Forward
glnAF: TGATAGGMACTTGGCAYCATATYAC
Reverse
glnAR: ARRCTCATATGMACATGCATACCA

glyA
Forward
glyAF: ATTCAGGTTCTCAAGCTAATCAAGG
Reverse
glyAR: GCTAAATCYGCATCTTTKCCRCTAAA

pgi
Forward
pgiF2: TTTAGTGGGWATGGGTGGKTCAAGT
Reverse
pgiR3: TCTCTAGCACCAATGAGAGCTATGG

tkt
Forward
tktF1: GCAAAYTCAGGMCAYCCAGGTGC
Reverse
tktR: TTTTAATHAVHTCTTCRCCCAAAGGT

Reaction conditions - PCR

Denaturation: 94ºC 30 s
Annealing: 53ºC 30 s
Extension: 72ºC 2 minutes
30 cycles