Bacterial 16S rRNA gene amplicon sequencing of marine sediment sample: GS10_PC15_1000
DNA was extracted using the PowerLyzer PowerSoil DNA kit according to manufacturers protocols (MOBIO), except for the addition of polyadenosine and use of 0.5-0.7 g of sediment. DNA was amplified with PCR (with HotStarTaq DNA Polymerase (QIAGEN)) using the bacterial-specific primers S-D-0564-a-S-15 (AYTGGGYDTAAAGNG) and S-D-Bact-1061-a-A-17 (CRRCACGAGCTGACGAC). Three replicates were pooled and purified using magnetic Agencourt AMPure XP beads (Beckman Coulter). Sequencing was performed with Illumina MiSeq (2x300 bp), with libraries constructed using adaptor sequences and reaction conditions from the TruSeq DNA LT Sample Prep Kit (Illumina).