DNA-Seq of Nephila clavipes: short insert (180 bp) fragment library of genomic DNA
Short insert, paired-end DNA fragment libraries were constructed using TruSeq LT kits following the "LS" protocol, revision C (Illumina). Genomic DNA was sheared to 300-600 bp using a Covaris S2 ultra-sonicator with clear microTUBEs, then end-repaired, A-tailed, ligated to adapters, purified, and size-selected at ~400 bp (180 bp insert + adapters) via gel electrophoresis (BioRad Low Range Ultra agarose). Gel slices were divided into 400 mg sections, processed using MinElute gel extraction kits (Qiagen), then PCR amplified as separate reactions in BioRad T100 thermocylcers using IlluminaŐs PCR Master Mix and PCR Primer Cocktail at recommended recipe volumes and standard cycling parameters. Final fragment libraries were cleaned using AMPure XP beads at a 1:1 ratio.