Amplicon Sequencing of Marine Sediment: Eastern Amundsen Sea Event 26  Core 4
The top 0-3 cm of marine sediment collected with a multicore and placed in a Whirl-Pak and frozen immediately in -80 freezer to be used DNA isolation for community composition analysis. Sediment was shipped back to Auburn University, Alabama, United States on dry ice and stored at -80C until extraction. Sediment was decanted over a 500 m sieve material retained on the sieve was saved. DNA was extracted from ~500 L of the retained slurry. Whole genomic DNA was sent to the Genomics Services Laboratory (GSL) at HudsonAlpha Institute of Biotechnology (Huntsville, AL) for metabarcoding sequencing of the hypervariable V9 region of the 18S small subunit ribosomal RNA (SSU rRNA) gene using eukaryotic specific primers 1380F and 1510R. Each sample was amplified in duplicate, and the duplicate reactions served as technical replicates. Each technical replicate had an initial input of 20 ng of DNA and remained as separate samples through sequencing and operational taxonomic unit (OTU) clustering. Reactions were dual-barcoded, pooled, and paired-end 100 bp reads were obtained by sequencing samples on an Illumina HiSeq 2500 platform.