Incubation of marine methane seep sediments with a bioorthogonal amino acid

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.