title: GSM2857834: MOLT16 input; Homo sapiens; ChIP-Seq
library_construction_protocol: 2x10^7 cells crosslinked with methanol-free formaldehyde (1% final concentration) RT for 7 min, reaction quenched with glycine . Nuclei isolation performed using truChIP Chromatin Shearing Reagent KIT (Covaris, PN520154). Purified chromatin fragmented by sonication (Covaris, M220, Focused-ultrasonicator). Chromatin fraction ON IP incubation with 40 μl of Protein A UltraLink™ Resin (Thermo-Scientific, 53139) and 4 μg Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (Abcam, ab4729). Day after, bead wash and ON reverse cross-linking by incubation at 65 °C. Input sample was also included (from 1.6x10^6 cells). DNA purified by Chromatin IP DNA Purification Kit (Active-Motif, 58002). library prepared using NEBNext Ultra DNA library Prep Kit (New England Biolabs,E73370S) and sequenced by NextSeq 500 (Illumina). DNA fragmentation, End repair, 5' Phosphorilation, dA-Tailing, Adaptor Ligation with U Excision, Size selection/CleanUp, PCR enrichment, Final CleanUp
attributes.tag: GEO Accession
attributes.value: GSM2857834