title: Metagenome derived from submerged sediments in Silver Bow Creek
design: Sediment samples dedicated for DNA analysis were collected concurrently with geochemical samples using an HDPE scoop, homogenized in a sterile specimen cup, partitioned into 2 mL autoclaved plastic tubes, frozen on dry ice in the field, and stored at 80 C until DNA extraction. DNA was extracted from collected sediments using a modified freeze-thaw and phenol-chloroform extraction procedure. Briefly, the sediments were mixed with DNA extraction buffer, homogenized, and the cells lysed using three successive freeze-thaw cycles. The sediment was then treated with a lysosome solution, proteinase K, and 20% sodium dodecyl sulfate (SDS) solution. Genomic DNA was then purified using a phenol/chloroform/isoamyl alcohol (24:24:1) solution, precipitated, treated with SEWS-M Wash Solution (MP Biomedicals), eluted, and cleaned using MOBIO PowerClean DNA Clean-Up Kit to a final extraction volume of 70 l. Universal bacterial primers 1100F and 1492R were then used to amplify the 16S rRNA genes present in the extracted DNA confirming that PCR-amplifiable DNA was present and discarded.The DNA from each sample was then sequenced at the University of Minnesota Genomics Center using the Illumina HiSeq 2500 platform with 2x100 bp sequences.
library_name: 151114F Storm Drain