16S rRNA gene amplicon sequencing of Bottle 5: maintained under strictly anaerobic conditions, 273 days of incubation
The cell pellet (10,000 x g centrifugation, 15 min) from 1 mL culture was frozen at -80 C for later DNA extraction using the Dneasy PowerSoil Kit (Qiagen) according to the manufacturer's procedure. Amplification and Illumina sequencing (MiSeq 300PE; paired-end) of extracted gDNA was carried out by McGill University at the Genome Quebec Innovation Centre. All amplicon sequences reads were generated using modified, staggered end primers 926F (AAACTYAAAKGAATWGRCGG) and 1392R (ACGGGCGGTGWGTRC), where 0- 3 random bases were inserted between both primer and Illumina adaptor sequences.