title: RAD-Seq of Liparidae
design: All sequences were generated from samples taken from whole specimens from the eastern North Pacific archived at the University of Washington. DNA was extracted using Qiagen DNeasy blood and tissue kit in single column format. DNA was concentrated to 6.25 ng/l in a 20 l volume using a vacuum concentrator and then quantified using the Quant-iT PicoGreen dsDNA Assay Kit and GEN5 software. Preparation of pooled RAD-tagged fragments was performed following the methods of Baird et al. (2008) and Hohenlohe et al. (2011). Following DNA digestion with SbfI, P1 adapters were ligated and a unique 6-nucleotide barcode was assigned to each individual pooled into a single library. DNA was randomly sheared to a target size of 300-500 bp using a Qsonica Q800R DNA sonicator, and size selected using Agencourt AMPure XP magnetic beads. A second set of adapters (P2) was added using blunt-end ligation. The base-pair target length of libraries (200400 bp), concentration, and quality were assessed with a 2100 Bioanalyzer and DNA 1000 kit. Single-read next-generation sequencing was performed as a single library in one direction at 150 bp target length (SR150) on a HiSeq 4000 at the University of Oregon Genomics Core Facility.
library_name: Cmelanu88