Gut associated bacterial community of cigarette beetle
A total ~300 ng from each amplicon DNA was end-repaired cleaned up with 1x AmPure beads. Barcoding adapter ligation (BCA) was performed with NEB blunt/TA ligase and cleaned with 1x AmPure beads.  Qubit quantified adapter ligated DNA samples were barcoded using PCR reactionand cleaned up with 1.6x AmPure beads.Qubit quantified barcode ligated DNA samples were pooled at equimolar concentration and end-repair was performed using NEBnext ultra II end repair kit. End-repaired DNA was cleaned up with 1x AmPure beads. Adapter ligation (AMX) was performed for 15 minutes using NEB blunt/ TA ligase. Library mix was cleaned up using Ampure beads and finally eluted in 15 l of elution buffer. Sequencing was performed on GridION X5 using SpotON flow cell R9.4 in a 48 hrs sequencing protocol. Nanopore raw reads (fast5 format) were base-called (fastq5 format) and de-multiplexed using Guppy tool.