Primers used for MLST of Staphylococcus lugdunensis
Genes
The Staphylococcus lugdunensis MLST scheme uses internal fragments of the following housekeeping genes:
aroE (shikimate dehydrogenase)
dat (D-amino acid aminotransferase)
ddl (D-alanine:D-alanine ligase)
gmk (guanylate kinase)
ldh (L-lactate dehydrogenase)
recA (recombinase)
yqiL (acetyl-CoA acetyltransferase)
Primers for PCR amplification and sequencing
| Gene | Direction | Primer Sequence |
|---|---|---|
| aroE | F | ATCGGAGATCCGATTTCACATTC |
| R | GGCGTTGTATTAATTATAATATC | |
| dat | F | TCGTGGTTATGTTTTTGGTGACGGT |
| R | ACAGTGTCTCCGCGATGCTGA* | |
| ddl | F | AGTGCGGAGCACGACGTTTCA |
| R | ACACTTGATCCCAAATTCGCCGGT | |
| gmk | F | ATAGTTCTTTCCGGACCATC |
| R | TCATTGACTACAACGTAATCATA | |
| ldh | F | ACTTGCAGGTGCCACGTCGA |
| R | GTTAGGAGAATTAGTTATGAAAGC** | |
| recA | F | GCACGGCCACCAGGTGTTGT |
| R | TCGGTAAAGGTGCCGTAATGAA*** | |
| yqiL | F | GTGCTAAACGCACACCAATTGGA |
| R | CTTCAGCATCGATTAGAGGCAC |
*(old primer, revoked on Feb 23rd, 2017: CTATGAGAAGTAAAGCCAGGAAT)
**(old primer, revoked on Feb 23rd, 2017: GCTACGCATTTGCAATGGTAACGCA)
***(old primer, revoked on Feb 23rd, 2017: AGGCCGTCGCGTATCTAGTGT)
PCR conditions
The PCR mixtures were heated for 3 min at 95°C, then a touch-down procedure followed, consisting of 30 s at 95°C, annealing for 30 s at temperatures decreasing from 60°C to 50°C during the first 11 cycles (with 1°C decremental steps in cycles 1 to 11), and ending with an extension step at 72°C for 30 s. A total of 40 cycles were performed.
Allele templates
Please refer to allele 1 from the sequence download page