GSM1402807: Wt1-2; Mus musculus; ChIP-Seq
On day E18.5 pregnant females were sacrificed by cervical dislocation and kidneys from embryos dissected and diced in ice cold Hanks Buffered saline solution. After fixation tissue was disrupted and nuclei released by homogenization in a 15ml Potter S homogenizer with Teflon plunger (B.Braun) attached to a drill in 20mM HEPES pH7.4, 1mM EDTA, 150mM NaCl, 1%SDS, 0.125M glycine, 0.2mg/ml PMSF. Nuclei were collected by centrifugation (805g for 5min) and re-suspended for sonication in 20mM HEPES pH7.4, 1mM EDTA, 150mM NaCl, 0.4%SDS, 0.2mg/ml PMSF. For library preparation, recovered DNA was digested with either AluI/MseI or HaeIII/MseI. Ends were filled and fragments A-tailed by treatment with GoTaq (Promega). Forked adapters (Illumina genomic DNA sample prep kit) were ligated to fragments and the resulting product was subject to 18 rounds of amplification using Illumina PCR primers and Pfx DNA polymerase (Invitrogen).
GEO Accession
GSM1402807