Whole genome bisulphite sequencing of Stylophora pistillata: 7.8B
Bisulphite DNA libraries were prepared following a modified version of the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, Ipswich, MA). Methylated TruSeq Illumina adapters (Illumina, San Diego, CA) were used during the adapter ligation step followed by bisulphite conversion with the EpiTect Bisulfite kit (Qiagen, Hilden, Germany), with the following cycling conditions (95 °C for 5 min, 60 °C for 25 min, 95 °C for 5 min, 60 °C for 85 min, 95 °C for 5 min, 60 °C for 175 min, then 3 cycles of 95 °C for 5 min and 60 °C for 180 min. Hold at 20 °C ? 5 hours). The final library was enriched with the KAPA HiFi HotStart Uracil+ ReadyMix (2X) (KAPA Biosystems, Wilmington, MA) following the standard protocol for bisulfite-converted NGS library amplification. Final libraries were quality checked using the Bioanalyzer DNA 1K chip (Agilent, Santa Clara, CA), and quantified using Qubit 2.0 (Thermo Fisher Scientific, Waltham, MA), then pooled in equimolar ratios and sequenced.
SUB2676380
7.8B_wgbs