title: SEDIMENT MICROBIOME
design: DNA was extracted  from sediment samples using Nucleospin soil kit (Macherey-Nagel, Germany) with minor modifications. The extracted DNA  was quantified using NanoDrop (Thermo Scientific, USA). A region of the 16S  rRNA gene covering the hyper variable V3-V4 was amplified by PCR using  bacterial-specific 341F-(5' CCTACGGGAGGCAGCAG 3) and 806R-(5'  GACTACHVGGGTATCTAATCC 3). The PCR products were sent to Scigenom Pvt Ltd  Kerala, India. The amplicon sequencing libraries were all prepared and  sequenced at Scigenom Pvt Ltd. Briefly, Amplicons of the correct size were  purified using PureLink Quick Gel Extraction and PCR Purification Combo Kit  (Invitrogen) following manufacturers instructions. A second PCR was performed  with primers that has Illumina indexed bar code sequences. The size selection  was performed with Pure link Gel extraction kit (Invitrogen). Purified  products were quantified using a Qubit (Invitrogen) The library preparation  was done using NEBNextUltra for DNA Library Prep Kit for Illumina. Sequencing  was done in MiSeq Illumina, 2 x 250bp  cycles. Sequence data was processed using QIIME.
library_name: SED S2