title: 16S rRNA gene sequencing of manganox enrichment: Santa Barbara enrichment Mn112 in defined freshwater medium with 1mM nitrate at 28.5C
design: For 16S rRNA gene amplicon sequencing, the V4-V5 region of the 16S rRNA gene was amplified from the DNA extracts using archaeal/bacterial primers with Illumina (San Diego, CA, USA) adapters on 5 end (515F 5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTGYCAGCMGCCGCGGTAA-3 and 926R 5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-CCGYCAATTYMTTTRAGTTT-3). Duplicate PCR reactions were pooled, barcoded, purified, quantified and sequenced on Illuminas MiSeq platform with 250 bp paired end sequencing. Raw reads with >1 bp mismatch to the expected barcodes were discarded, and indexes and adapters were removed using MiSeq Recorder software (Illumina).