title: GSM5237549: nrpd1-br1 (methylation); Arabidopsis thaliana; Bisulfite-Seq
library_construction_protocol: Genomic DNA was isolated using the Dneasy Plant Mini Kit (Qiagen) 5 ug of genomic DNA was used to generate the MethylC-seq libraries as described by Lister et al. 2008 with minor modifications. genomic DNA was fragmented to approximately 200 bp by sonication with a Covaris sonicator according to manufacturer's instructions (Covaris, S220), then subjected to end repair and ligation of methylated adapters provided by Illumina (Illumina, FC-121-2001) per manufacturer's instructions for gDNA library construction. 100-200 ng of adapter-ligated gDNA was subjected to sodium bisulfite treatment using the MethylCode Bisulfite conversion kit (Thermo Fisher, MECOV50). Converted and adapter-ligated DNA fragments were enriched by 12 cycles of PCR amplification with PfuTurboCx Hotstart DNA polymerase (Thermo Fisher) . The library was purified with the PCR purification kit (Invitrogen, K310001) and quantified on a Bioanalyzer Instrument (Agilent).