Sequencing of protein-coding genes is an increasingly used technique in microbiology laboratories. The analysis of these sequences enables us to establish phylogenetic relationships and improve species-level identification in those species for which ribosomal DNA analysis does not provide sufficient resolution.

The choice of genes depends on the microbial group to which the isolate belongs. Some of the most commonly used in prokaryotes are: recA, gyrB, ftsZ, rpoD, mreB.

Necessary material

Preferably, an active culture of the strain (petri dish or agar slant).

If the customer prefers, we can also work with genomic DNA of the strain. In that case, the genomic DNA must be delivered in cold-storage and be of good quality (A260 / A280 between 1.8 and 2). We require more than 1 microgram of DNA at a concentration of 40-50 ng/µl resuspended in ultrapure water or in TE buffer.

Methodology

Should the customer decide which genes are to be sequenced, a citation must be supplied with the study methodology (sequence of primers, PCR conditions, etc.).

Otherwise, the CECT staff will perform a literature search and advise the client on which gene / genes may be useful for the strain under study.

Sequencing is done following the methodology described in the literature for the chosen gene.