16s rRNA Metagenomics of Mangrove rhizosphere: Madhwad region
Genomic DNA was extracted using Qiagens DNeasy PowerSoil DNA isolation kit using homogenized soil in duplication. For studying bacterial diversity, V6-V8 region of 16s rRNA, which is around 400bp, was amplified using Phusion forward (CGATACTGGCGGACGGGTGAGTAA) and reverse (ATCGTATTACCGCGGCTGCTGG) primers which contained adapter tag and a unique barcode. PCR products were purified by using the QIAquick Gel Extraction kit. Library was prepared using Qiagen OT2 kit as per manufacturers protocol. Amplicon library was quantified using Qubit Flurometer 2.0 (Life Technologies). Sequencing was done using Ion 316TM Chip v2 BC using Ion PGMTM Hi-QTM View Sequencing Kit in Ion PGM sequencer as per manufacturers protocol.