⚠ Maintenance: A BIGSdb-Pasteur maintenance is scheduled on Tuesday, May 12th (Central European Time). During this period, the server and API will be unavailable. Please note that job submission will not be available and all unfinished jobs will be killed. Sorry for any inconvenience. If any questions, contact us
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