alias: GSM3678308
title: GSM3678308: iSTAT3β_180-min-OSM_ChIPSeq; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were rinsed with PBS and cross-linked with 1% (w/v) formaldehyde at room temperature (10 min). The cross-linking reaction was quenched by adding 0.1 volume of 1.25 M glycine and incubated at room temperature (5 min). The cells were then rinsed with cold PBS and lysed in 5 mL nucleus isolation buffer (20 mM Tris-HCl pH 8.0, 85 mM KCl, 0.5% (v/v) NP-40) supplemented with Complete™ protease inhibitor mix (Roche Diagnostic, USA). The pellet, containing the crude nuclear preparation, was collected after centrifugation (1,200 × g, 5 min, 4°C) then lysed in 1 mL cold nucleus lysis buffer (150 mM NaCl, 50 mM Tris-HCl pH 7.3, 1 mM EDTA pH 8.0, 0.5% (w/v) sodium deoxycholate, 1% (w/v) SDS, 1% (v/v) NP-40) supplemented with Complete™ protease inhibitor mix.  The resulting nuclear extract was then sonicated (4°C, high setting, 20 cycles: 30 seconds on, 30 seconds off) in a Biorupter® Pico (Diagenode, USA) to obtain 200-500 bp DNA fragments. Sequencing libraries were constructed using the NEBNext Ultra II DNA Library Prep kit (New England Biolabs, USA) according to the manufacturer's protocol. Libraries were sequenced using the Illumina NextSeq 550 System (Illumina, USA). Duplicate samples were prepared and analysed.
identifiers.namespace: GEO
identifiers.id: GSM3678308