alias: S-linearis-S6
title: Low coverage genome sequencing of the tamarind weevil Sitophilus linearis
design: Twenty pairs of ovaries were dissected from females S. oryzae, S. zeamais, S. granarius and S. linearis. Ovaries were homogenized in 100 mM NaCl, 1 mM EDTA pH 8, 10 mM Tris-HCl pH 8 using a small piston. Proteinase K digestion followed in the presence of SDS for 2h at 55C with shaking and for 1h at 37C with RNAse A. A typical phenol chloroform extraction was then performed and genomic DNA was isopropanol precipitated. Eight whole genome sequencing libraries with a median insert size of 550 bp were constructed using the Illumina TruSeq DNA PCR-free sample preparation kit, according to manufacturer's protocols. Briefly, 2 ug of each gDNA were sheared using a Covaris M220 Focused-ultrasonicator, end-repaired, A-tailed, and adapter ligated. Library quality control was performed using the 2100 Bioanalyzer System with the Agilent High Sensitivity DNA Kit. The libraries were individually quantified via qPCR using a KAPA Library Quantification Kits for Illumina platforms, then they were pooled together in equimolar quantities and sequenced in a MiSeq sequencing system (Illumina). 2x300 paired-end reads were obtained using a MiSeq Reagent Kits (600-cycles).
identifiers.namespace: SUB7790883
identifiers.id: S-linearis-S6