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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
| Gene | Direction | Sequence |
|---|---|---|
| abcZ | abcZ-F | TGACGACCAAGCGAGCGTGTTTGA |
| abcZ-R | TCCGCCTCAACCGCCAATTCCT | |
| adk | adk-F | CACAAGCGCAATTTATTACGCGCGA |
| adk-R | GTCGTCATCGCGTTGCACCAAGT | |
| aroE | aroE-F | TTTGCCGCACAAGAAGGTGCGCAA |
| aroE-R | GGCTGGTTCGCATAAAACATATCG | |
| cpn60 | cpn60-F | TGTTGGCGCAAGCGATTGTTGCT |
| cpn60-F-3R | AAACCAATGATGTGGCTGGCGACG* | |
| cpn60-R | AATGGGCTGTCCAAACCAGCGAT | |
| gdh/zwf** | gdh-zwf-F | AGGCGCATTGCGCGACATGATGCA |
| gdh-zwf-R | CCAGTTGTCCAAAATTGGCATGAC | |
| recA | recA-F | GACGAAGAATTGCAAGTCATTTCCA |
| recA-R | AGTTTACGCAAGGCTTGGCTCATC |
*(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