title: GSM2400009: cR-S6EOS-2_P5#1; Homo sapiens; Bisulfite-Seq
library_construction_protocol: Post-bisulfite adaptor tagging (PBAT) libraries for whole-genome DNA methylation analysis were prepared directly from cell lysates following recently described protocols (Miura et al., 2012; Smallwood et al., 2014). PBAT libraries were prepared by bisulfite converting Proteinase K digested cell lysates using the EZ DNA Methylation-Direct Kit (Zymo). After cleanup, 1st strand synthesis was performed using Klenow Exo- DNA Polymerase (NEB) and 6N-forward oligos at 37°C for 90 minutes. Subsequently, samples were treated with Exonuclease I (NEB) for 1 hour at 37°C, and DNA was purified using AMPure XP beads (Beckman Coulter). Samples were eluted in 2nd strand synthesis mix with Klenow Exo- and 6N-reverse oligos and incubated at 37°C for 90 minutes. DNA was purified and amplified with KAPA HiFi HotStart DNA Polymerase (Roche). All amplified libraries were purified and assessed for quality and quantity using High-Sensitivity DNA chips on the Agilent Bioanalyzer. High-throughput sequencing was carried out with a 125 bp paired-end protocol on a HiSeq 2000 instrument (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM2400009