Shotgun metagenomics sequencing of stool DNA
Genomic DNA were end-repaired, cleaned up with 1X AmPure beads. Adapter ligation was performed with NEB blunt/ TA ligase and cleaned with 0.8X AmPure beads. Adapter ligated DNA samples were barcoded using PCR reaction, cleaned up with 1.6x AmPure beads. Barcode ligated DNA samples were pooled at equimolar concentration and end-repair was performed. End-repaired DNA was cleaned up with 1x AmPure beads. Adapter ligation 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.