Hot spring enrichment culture 16S rRNA gene tag sequencing

Microbial members of high-temperature geothermal springs are relatively understudied in cultivation-dependent conditions. To establish stable cultivation of these thermophilic archaeal and bacterial lineages, sediment from Great Boiling Spring was used to inoculate media containing 22 different substrate and growth conditions to include various organic carbon sources, electron acceptors, oxygen concentrations, and temperatures. Sequence-based community composition of each enrichment was measured at two weeks of incubation after primary inoculation and at four weeks of incubation after first transfer.