16S rRNA gene amplicons from iron addition experiments in mesoscale eddies: DCM control from cyclonic eddy
Natural seawater was incubated at collection depth (with or without Fe addition) on an in situ array in 2L bottles for 64 hours, then cells were collected on 0.2 m polyethersulfone filters, preserved with RNA-later, and frozen at -80. Cells were lysed by freeze-thawing and bead-beating, then genomic DNA was extracted using the MasterPure Complete DNA and RNA Purification Kit (Lucigen). PCR was conducted using primers 515F-Y and 926R as recommended by Parada et al. 2016, with multiplexing indexes as designed by the Earth Microbiome Project. Triplicate amplifications were cleaned using an ENZA Cycle Pure Kit (Omega Bio-tek) and pooled at equimolar proportions. The pooled library was sequenced on an Illumina MiSeq using PE250 v2 chemistry at the University of Montana Genomics Core.