Hydroides elegans genomic DNA 300bp
DNA was extracted from a single male worm using a Qiagen DNeasy blood and tissue kit according to the manufacturer’s instructions. Two libraries with average insert sizes of 300 bp and 600 bp were prepared using NEBNext Ultra DNA library preparation kit (NEB #E7370) according to manufacturer’s instructions with minor modifications. For each library, 1 μg of genomic DNA was fragmented using Qsonica Q800R sonicator to generate the target size distribution. The fragmented DNA underwent end repair and dA tailing followed by adaptor ligation. Adaptor ligated DNA was size selected on agarose gel and sequences required for multiplexing and clustering were added by 6 cycles of PCR. PCR products were purified with Agencourt AMPure XP beads (Beckman Coulter). Libraries were quantified using Qubit fluorometer and the dsDNA HS Assay Kit (Invitrogen) and the size distribution was assessed with 2100 BioAnalyzer and the High Sensitivity DNA Kit (Agilent).