Metabarcoding approch is conventionally based on total gDNA isolation present in a given sample, making not possible the differenciation between living and active bacteria, from dead and non-functional bacteria that released their DNA in the sample.
Illumina “short read” sequencing limits the sequencing of short fragments (maximum 550 bp), which allows even the sequencing of degraded gDNA of dead bacteria.
It thus comes to the limit of this 16S rRNA sequencing method from gDNA, for the distinction of living from dead bacteria.
In collaboration with the “ Staphylococcal pathogenesis” team of the International Center for Infectiology Research of Lyon (INSERM U1111 – CNRS UMR5308 – ENS Lyon – Lyon 1 University), more particularly with Dr Jean-Philippe RASIGADE and Anaïs BARRAY, we have developed a method allowing sequencing and identification only of live and active bacteria in a given sample.
If you are interested in this approach, please do not hesitate to contact us.