GSM5026063: BSseq_lib1_mbd56_rep2; Arabidopsis thaliana; Bisulfite-Seq
DNA was extracted with DNeasy Plant Mini kit (Qiagen) 200ng of DNA per sample were sheared with the Covaris S2 instrument to an average fragment size of 200bp (Duty Cycle = 10%, Intensity = 5, Cycles per Burst = 200, Treatment Time = 120 seconds). A-tailing and adapter ligation was performed with the Kapa Hyper Prep Kit (KR0961) using Illumina methylated adapters (TruSeq DNA Sgl index Set A or B). Bisulfite conversion of the DNA was carried on with the EpiTect Bisulfite Kit (Qiagen). The conversion reaction in the thermocycler was performed twice to increase the efficiency. The converted library was PCR amplified using myTaq HS mix (Bioline) with 12 PCR cycles, purified with Ampure XP beads (Beckman Coulter) and sequenced.