title: GSM4801065: MeDIP_P1_rep3; Mus musculus; MeDIP-Seq
library_construction_protocol: DNA was extracted from 500,000 - 1 million mES cells using phenol-chloroform-isoamyl alcohol (Thermo) extraction. 500ng of purified DNA was subjected to sonication on a Covaris S220 sonicator (10% duty cycle, Intensity 5, 200 cycles/burst, 6 one minute cycles). After sonication fragmented DNA was end repaired and A-tailed using Ultra II End Repair/dA-Tailing Module (NEB) then adapters (Broad Institute, single index P7) were ligated using Blunt/TA Ligase Master Mix (NEB). Both protocols were carried out as recommended by the manufacturer. MagMeDIP kit (Diagenode) was used for precipitation and wash steps according to manufacturer's recommendations. Libraries were PCR amplified with universal primers to add the P7/P5 graft sites using Phusion High-Fidelity PCR Master Mix with HF Buffer (NEB) then size selected between 200-700bp on an E-Gel Agarose Gels 2% (Thermo). DNA was purified using MinElute Gel Extraction Kit (Qiagen). Purified DNA was quantified using Bioanalyzer HS DNA (Agilent) and qBit HS dsDNA (Thermo) kits then sequenced on a HiSeq 4000 system. 
attributes.tag: GEO Accession
attributes.value: GSM4801065