GSM2807190: WT_bsseq; Arabidopsis thaliana; Bisulfite-Seq
Genomic DNA (gDNA) was extracted from 1-month-old Arabidopsis thaliana rosette leaves with the DNeasy plant mini kit (Qiagen, cat. no. 69104) per the manufacturers' instructions. Libraries were prepared from roughly 500ng of purified gDNA that was sheared to approximately 400bp on a Misonix water bath sonicator, then purified using 1.2X volume of Agencourt Ampure beads (referred to as “beads” henceforth, cat. no. A63881). Following ligation of methylated Truseq sequencing adapters (Illumina), bisulfite conversion of DNA was carried out according to manufacturer's protocol (Qiagen Epitect Kit, cat. no. 59104) except without using carrier RNA. DNA was purified twice with 1.2X beads and converted a second time to ensure complete bisulfite conversion of unmethylated cytosine. NEB next indexing primers (cat. no. E7335S) were used for generating multiplexed libraries during PCR amplification of libraries.
GEO Accession
GSM2807190