16S rRNA gene sequencing of: Monterey Canyon, cold seep SW Clam Patch 1, sediment 18-21 cm
DNA was extracted with the DNeasy PowerSoil Pro Kit (Qiagen 12888; Hilden, Germany) according to the kit protocol with an added bead beating step. The V4-V5 region of the 16S rRNA gene was PCR amplified in duplicate using archaeal/bacterial primers 515f (5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTGYCAGCMGCCGCGGTAA-3) and 926r (5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGCCGYCAATTYMTTTRAGTTT-3). Duplicates were pooled and amplified with Illumina Nextera XT index 2 primer barcodes. Equimolar amounts of barcoded samples were combined, purified, and submitted to Laragen (Culver City, CA) for PE250 sequencing on Illuminas MiSeq platform with addition of 15-20% PhiX control library.
SUB16166911
MontereyCanyon_SWclams1_18_21_cm_16S