Primers used for MLST of Kingella kingae

Genes

The Kingella kingae MLST scheme uses internal fragments of the following housekeeping genes:

abcZ (ABC transporter)
adk (adenosine kinase)
aroE (shikimate dehydrogenase)
cpn60 (heat shock protein)
gdh/zwf (glucose-6-phosphate 1-dehydrogenase)
recA (DNA repair and recombination)


Primers for PCR amplification and sequencing

GeneDirectionSequence
abcZabcZ-FTGACGACCAAGCGAGCGTGTTTGA
abcZ-RTCCGCCTCAACCGCCAATTCCT
adkadk-FCACAAGCGCAATTTATTACGCGCGA
adk-RGTCGTCATCGCGTTGCACCAAGT
aroEaroE-FTTTGCCGCACAAGAAGGTGCGCAA
aroE-RGGCTGGTTCGCATAAAACATATCG
cpn60cpn60-FTGTTGGCGCAAGCGATTGTTGCT
cpn60-F-3RAAACCAATGATGTGGCTGGCGACG*
cpn60-RAATGGGCTGTCCAAACCAGCGAT
gdh/zwf**gdh-zwf-FAGGCGCATTGCGCGACATGATGCA
gdh-zwf-RCCAGTTGTCCAAAATTGGCATGAC
recArecA-FGACGAAGAATTGCAAGTCATTTCCA
recA-RAGTTTACGCAAGGCTTGGCTCATC

*(For a few strains, in case of failure with cpn60-F)
**(Sequences of gdh primers were redefined on August 30th, 2016 based on whole genome sequencing)


PCR conditions

Conditions for PCR amplification are 15 minutes at 95°C followed by 35 cycles of 95°C for 15 s, 56°C for 30 s and 72°C for 1 min 30 s with a final extension step at 72°C for 10 minutes.


Allele templates

Please refer to allele 1 from the sequence download page

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