GSM3465883: h1_bisulfite_rep2; Arabidopsis thaliana; Bisulfite-Seq
gDNA for bisulfite sequencing was extracted with Dneasy plant mini kit according to manufacturer's protocol. To construct bisulfite sequencing libraries, about 500 ng of purified gDNA was fragmented to roughly 100-1000 bp then purified using 1.2X volume of SPRI beads (Beckman Coulter, cat. No. A63881). Fragmented gDNA was end-repaired and ligated to HPLC/PAGE purified, cytosine-methylated Illumina adapters (Elim Biopharmaceuticals, Inc.). DNA was subjected to two rounds of bisulfite conversion according to manufacturer's protocol (Qiagen, cat. No. 59104). To remove self-ligated adapters, DNA was purified twice with 0.8X volume of SPRI beads. NEB next indexing primers were used to amplify bisulfite-converted libraries (New England BIolabs Inc. cat. No. E7335S). DNA fragments isolated for ChIP-, ChAP-, and DnaseI-sequencing were subjected to Illumina library construction as described in bisulfite sequencing library construction except for the two rounds of bisulfite conversion.
GEO Accession
GSM3465883