alias: GSM3375774
title: GSM3375774: LRH-1_+OvS; Mus musculus; ChIP-Seq
library_construction_protocol: Ovarian tissues collected following 0 and 4h post hCG (n=5 per condition) were washed in PBS, sliced in small pieces, and fixed 15 min in 1X PBS/1.1% formaldehyde with agitation. After glycine add and PBS wash, fixed tissues were incubated in lysis buffer and mechanically disrupted with an ice-cold Dounce. Chromatin was then sonicated in a Bioruptor apparatus (Diagenode) on “high”-intensity position for 30 s ON and 30 s OFF for 15 cycles at 4 °C, cleared by a 10 minutes centrifugation at 10 000 x g and checked on agarose gel. ChIP experiments were performed using 10µg of LRH-1 antibody with the use of magnetic protein A dynabeads (Life Technologies) for the immunoprecipitation. After wash, elution and decross-link, samples were purified using QIAquick® PCR purification kit (QIAGEN) and quantified with PicoGreen™ assay (Invitrogen). Librairies were performed as previously described (Bianco et al., Methods Mol Biol, 2015)
identifiers.namespace: GEO
identifiers.id: GSM3375774