GSM3319581: drm1/drm2; Physcomitrella patens; Bisulfite-Seq
Genomic DNA was isolated from protonema, fragmented by sonication, end repaired, and ligated to custom synthesized methylated adaptors. Adaptor-ligated libraries were subjected to two successive treatments of sodium bisulfite conversion using the EpiTect Bisulfite kit (QIAGEN) as outlined in the manufacturer's instructions. The bisulfite-converted libraries were then amplified by PCR and either gel purified (~300 bp band) or purified with the solid-phase reversible immobilization method using AM-Pure beads (Beckman Coulter) prior to quantification with a Bioanalyzer (Agilent). DNA was extracted from approximately 500 mg tissue using NucleoSpin® Plant II Midi kit for Genomic DNA extraction from plant and fungi (Macherey-Nagel), according to recommended protocol