title: GSM4830453: 2047_HV [RRBS]; Homo sapiens; Bisulfite-Seq
library_construction_protocol: Genomic DNA was isolated using the AllPrep DNA/RNA mini kit according to the manufacturer's instructions (Qiagen). Libraries were prepared from 100ng genomic DNA using Diagenode Premium RRBS kits (Diagenode). Briefly, DNA was digested with the restriction enzyme MspI, which recognizes CCGG dinucleotides sites, and AMPpure XP kit (Beckman Coulter) for size selection. Samples were pooled based on concentrations (n=8 per pool) and treated with sodium bisulfite to convert the unmethylated cytosines to uracil leaving unmethylated cytosines unchanged. After bisulfite conversion samples were amplified by PCR using the following reaction setup: 20 minutes at 25°C, 10 minutes at 65°C, hold at 8°C. PCR products were cleaned using the AMPpure XP kit and sequenced on the Illumina HiSeq4000 (Illumina) instrument in single read (50) mode.