title: GSM3543740: WSD24 (RRBS); Macaca mulatta; Bisulfite-Seq
library_construction_protocol: DNA extracted using the AllPrep DNA/RNA purification kit (QIAGEN) according to the manufacturer's instructions RRBS libraries were generated starting with genomic DNA extracted from WAT.  Overnight digestion was performed on ~200ng DNA using the restriction enzyme MspI (New England Biolabs, Ipswich, MA). Reaction clean-up was performed with AMPure XP magnetic beads (Beckman Coulter, Brea, CA) before use with the NEXTflex Bisulfite-Seq Kit (Bioo Scientific Corporation, Austin, TX) for library preparation.  The digested DNA was quantified and then ~80ng was end-repaired, A-tailed, and ligated with the NEBNext Methylated Adaptor (New England Biolabs).  The ligated DNA was cleaned and then size-selected using AMPure XP magnetic beads to produce a final library size of 350 bp.  Bisulfite conversion was performed with the EZ DNA Methylation-Gold Kit (Zymo Research, Irvine, CA) before carrying out PCR amplification with NEBNext Multiplex Oligos for Illumina (New England Biolabs) to barcode each library. Two AMPure XP bead cleans were performed and the resulting libraries were quantified with the Qubit High Sensitivity dsDNA Assay (Life Technologies, Eugene, OR) and the Bioanalyzer High Sensitivity DNA Kit (Agilent, Santa Clara, CA).
attributes.tag: GEO Accession
attributes.value: GSM3543740