Metagenomic analysis of two banana cultivars Red Decca and Ney Poovan - NP_Tr
A total of 50 ng of DNA from each sample was subjected to PCR amplification using barcoded 16S V1-V9 region specific primers (27F  AGAGTTTGATCMTGGCTCAG and 1492R  TACGGYTACCTTGTTACGACTT) and LongAmp (NEB) Taq 2X master mix, following the protocol outlined in the 16S Barcoding Kit 24 V14 (SQK-16S114.24; Oxford Nanopore). The amplicons contain 5 tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The amplicons generated from the samples were confirmed using 1% agarose gel electrophoresis stained with EtBr. The PCR products were purified by using 1.6X Ampure XP beads (Beckmann Coulter, USA). The amplicons from each sample were quantified using Qubit 4 Fluorometer (Thermo Fisher Scientific, USA) and ~50 ng from each sample were pooled. The pooled amplicons were subjected to library clean-up using the 1X AmPure XP beads (Beckmann Coulter, USA) supplied in the 16S Barcoding Kit 24 V14. Sequencing was performed on PromethION 2 Solo platform (Oxford Nanopore Technologies, Oxford, UK) using R10.4.1 flow cells (FLO-PRO114) on MinKNOW (version 24.06.14, ONT).