Amplicon Sequencing of Halichoerus grypus: Mitochondrial Control Region from Environmental Samples
DNA was extracted from water samples collected from gray seal haulout sites around Cape Cod, Massachusetts, USA. In addition to sequencing these samples, we also sequencced positive controls (diluted tissue extracts), negative PCR controls containing no DNA extract, and a series of mock communities comprised of diluted gray seal tissue extracts mixed at various concentrations. We amplified a 381-384 bp fragment of the mitochondrial control region using species-specific gray seal primers (Forward: 5-CTGACATCCACTCAACCCCC, Reverse: 5-TGGATTCTAGGACTTTCATGGC) and TruSeq Illumina adapters. PCR products were purified with Beckman Counter Agencourt AMPure XP beads at 1.8x concentration and sequenced  at the Hubbard Center for Genome Studies (University of New Hampshire, USA) on an Illumina NovaSeq (2x250bp). This dataset is comprised of three sequencing runs that were prepared with the same methodology but sequenced separately.