Incubation of marine methane seep sediments with a bioorthogonal amino acid

Sampling

In marine sediments, anaerobic oxidation of methane (AOM) is primarily catalyzed by syntrophic consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). We applied bioorthogonal non-canonical amino acid tagging (BONCAT) using a high-throughput approach to detecting protein synthesis in individual cells, on sediments from deep-sea methane seeps. Sediment samples have been incubated in the presence or absence of methane and a bioorthogonal amino acid. V6 regions of bacterial and archaeal 16S rRNA genes were amplified using general primers, tagged, and analyzed for shifts in the microbial community during time of incubation.

Method steps

1. Pipeline used: https://www.ebi.ac.uk/metagenomics/pipelines/4.1