title: DNA-seq of E. tetradactylum: Developing male fish muscle
design: The paired-end sequencing libraries were prepared from the QC passed gDNA samples (KET25; KET29 and KET30) using TruSeq Nano DNA library Prep Kit for illumina. 200ng of Genomic DNA from the three samples was fragmented by Covaris M220 to generate a mean fragment distribution of 400bp. Covaris shearing generates dsDNa fragments with 3 to 5 overhangs. The fragments were then subjected to end-repair. This process converts the overhangs resulting from fragmentation in to blunt ends using End Repair Mix. The 3 to 5 exonuclease activity of this mix removes the 3overhangs and the 5 to3 polymerase activity fills in the 5 overhangs followed by adapter ligation to the fragments. This strategy ensures a low rate of chimera formation. The ligated products were size selected using AMPure XP beads. The size selected product was PCR amplified with the index primer. Indexing adapters were ligated to the ends of the DNA fragments, preparing them for hybridization on to a flow cell.