design: The DNA fragments undergo end repair where in the mix converts the overhangs resulting from fragmentation into blunt ends. The 3 to 5 exonuclease activity of end repair mix removes the 3 overhangs and polymerase activity fills in the 5 overhangs. To the blunt ended fragments adenylation is performed by adding single A nucleotide to the 3 ends. To the adenylated fragments loop adapters were ligated and cleaved with uracil-specific excision reagent (USER) enzyme. The samples are further purified using AMPure beads and The DNA is then enriched by PCR with 6 cycles using NEBNext Ultra II Q5 master mix, illumina universal primer and sample-specific octamer primers.