Sediment 16S rDNA amplicon sequence metagenome
Genomic DNA was extracted in triplicate from the 0.25g sediment samples using Fecal/SoilTotal DNATM extraction kit (Zymo Research Corporation, CA, USA) according to manufacturers instructions and stored at 20C prior to further analysis.The quantity and the quality of the resultant genomic DNA was checked on a 1.5% agarose gel and Biodrop Lite spectrophotometer (Biochrom Ltd, Cambridge, UK). Extracted DNA used for downstream PCR and sequencing analysis had A260:A280 ratios between 1.82.0 and DNA concentrations of 20150 ng/L. 16S rRNA gene fragments libraries preparation using 27F (5-AGAGTTTGATCMTGGC-3) and 518R (5-GTATTACCGCGGCTGCTGG-3), fused with MiSeq adapters and heterogeneity spacers compatible with Illumina indexes for multiplex sequencing, was done according to protocol described by Ogola et al (2021). Resultant libraries were sequenced by paired end (300 bp reads) sequencing v.3 chemistry along with its multiplex sample identifiers on the Illumina MiSeq 250 Platform according to standard protocol.