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