title: GSM8484493: input-b-Catenin_FL_E10.5+DMSO+6h; Mus musculus; ChIP-Seq
library_construction_protocol: Dissected forelimb bud tissue was crosslinked in 1% formaldehyde/PBS at room temperature which was then quenched with glycine (125 mM). For β-Catenin ChIP a second crosslink step was performed using DSG (disuccinimidyl glutarate) for 40 min at RT. After lysing in hypertonic buffer chromatin fragments were generated by sonication. Immunoprecipitation was performed overnight at 4°C using the polyclonal anti beta Catenin antibody (71-2700 Invitrogen, 5 μg per sample) and anti-histone H3(acetyl K27) antibody (ChIP Grade ab4729 Abcam, 5 μg per sample). The immune-complexed chromatin complexes were extracted with magnetic beads (Fisher Scientific 11202D). After washing beads in RIPA buffer, DNA was eluted from beads followed by reverse cross-linking overnight. Libraries were generated using the next-generation library preparation kit from Takara Bio (Japan) according to manufacturer instructions ChIP-Seq libraries were sequenced using a NextSeq instrument (Illumina).