PacBio Smrt II WGS Bombus terricola gyne Bcola_28 part B thorax
High quality, high molecular weight genomic DNA was extracted from one Bombus terricola gyne using Qiagen Genomic-tip protocol and Qiagen Blood and Cell Culture DNA mini kit (Qiagen, Hilden, Germany) using manufacturer s protocol with modifications in the lysis step. Specifically, 20mg of bee tissue (comprising thorax and legs) was divided into two parts and lysis step was carried out in two tubes of ~10mg tissue  as starting material. For each part, the tissue was ground using liquid nitrogen, 350  l CTAB extraction buffer (0.1 M Tris-HCl at ph8.0, 1.4 M NaCl, 0.02 M EDTA, 2% Cetyltrimethyl ammonium bromide) and 2 l RNAse A added, and incubated at 37C for 30min. To this 20 l Proteinase K and 1ml Buffer G2 (supplied in the kit) was added and incubated overnight at 50C. Next morning, the lysates from two tubes were vortexed and added to one Genomic-tip column. The protocol from here onwards was followed as specified in the kit s manual. The size of the genomic DNA was evaluated on a pulse-field gel electrophoresis. The DNA was further quantified and checked for purity using Nanodrop. The high quality DNA isolated from this B. terricola gyne was sent to G nome Qu bec Innovation Centre (Montreal, Canada) for generating libraries and sequencing using the Pacific BioSciences RS II long read technology (Pacific Biosciences, Menlo Park, CA, USA). A total of 15 SMRT cells were used for this one sample to generate ~35X coverage.
SUB3342554
Pbio_Bcola_28B