Post _TSS
A total of 500ng of DNA from each sample was end-repaired (NEBnext ultra II end repair kit, New England Biolabs, MA, USA), cleaned up with 1X AmPure beads (BeckmannCoulter,USA). Barcode adapter ligation (BCA) was performed with NEB blunt/ TA ligase (New England Biolabs, MA, USA) and cleaned with 0.8X AmPure beads. Qubit quantified adapter ligated DNA samples were barcoded using PCR reaction (LongAmp Taq 2x New England Biolabs, MA, USA) and cleaned up with 1.6x AmPure beads (Beckmann-Coulter, USA). Barcode sequences are detailed in the table 1 below. Qubit quantified barcode ligated DNA samples were pooled at Equimolar concentration to attain 1 g pooled sample and end-repair was performed using NEBnext ultra II end repair kit (New England Biolabs, MA, USA). End-repaired DNA was cleaned up with 1x AmPure beads. Adapter ligations (AMX) was performed for 15 minutes using NEB blunt/ TA ligase (New England Biolabs, MA, USA). Library mix was cleaned up using Ampure beads and finally eluted in 15 l of elution buffer.