Dataset: Metagenomic, and single cell sequencing data from a Fall 2014 ESP deployment in Monterey Bay, CA

The Environmental Sample Processor (ESP) filtered seawater sequentially through 0.2 um pore size polyethersulfone filters. Seawater was evacuated from filters and followed twice with a 2 minute incubation with 1 ml of RNAlater™. RNAlater was evacuated, and filters were stored in the ESP until they were transferred to -80 C upon instrument recovery. 

Filters were processed for DNA using the phenol-chloroform extraction method of Crump et al. (2003) after placing the filters into 1 ml of DNA extraction buffer. Extracted DNA was sheared ultrasonically to ~350 bp fragments, and library preparation was performed at the Georgia Genomics and Bioinformatics Core (GGBC) facility. Single-end 250 bp sequencing was performed using an Illumina HiSeq Rapid Run at Hudson Alpha Genomic Services Laboratory (Huntsville, AL, USA).

Single-cell sequencing: Seawater was transferred directly from the Niskin bottle to a 50 ml Falcon tube and placed on ice until brought back to lab. Each sampling day, 3 x 1 ml of seawater was preserved in cryovials using 100 ul of glyTe (5 ml glycerol, 3 ml Milli-Q H2O, 1 ml 100 x TE pH 8.0, 0.2 um filter sterilized after mixing the above, and stored in -20 C freezer). Preserved samples were then placed in a -80 C freezer. Samples were processed and sequenced at Bigelow Single Cell Genomics Center (Stepanauskas and Sieracki, 2007).