16S rRNA library of seawater dilution experiments - Expt#1 Summer Day5 Initial A
DNA was extracted from half of each filter (cut with sterile razor blades) using the Gentra Puregene Yeast/Bacteria Kit (QIAFEN) with the addition of bead beating (with 0.1 mm zirconia/silica beads) for 3 cycles of 30 seconds. Extracted DNA was cleaned with the OneStep PCR inhibitor removal kit (Zymo), quantified by using DeNovix DS 11FX+, and diluted to ~ 20 ng/μL. 16S rRNA gene primers targeting the V4–V5 region 515F-Y (5′-GTGYCAGCMGCCGCGGTAA) and 926R (5′-CCGYCAATTYM TTTRAGTTT) with added barcodes and adapters were used to construct 16S rRNA gene libraries (Parada, Needham, and Fuhrman 2016).
SUB15390096
E1_SU_D5_I_A