title: GSM2974457: ChIP-seq of Mybl2 in MEF; Mus musculus; ChIP-Seq
library_construction_protocol: 2 x107 cells were suspended in 30 mls PBS and incubated with 250 ul DSG (50mg/500ul DMSO) on a rotating wheel for 45 minutes at room temperature. After this step, cells were washed 4 times with PBS and then resuspended in 10 ml of PBS and used for the second cross-linking with 1% Formaldehyde for 10 minutes at room temperature. Cross-linking was then terminated by adding 4 volumes of cold PBS+0.125 M of Glycine. Following this step cells were resuspended in 300l of ice-cold ChIP Lysis Buffer (50mM HEPES-KOH pH 7.5, 1mM EDTA pH 8, 1% Triton X-100, 0.1% Na Deoxycholate and proteinase inhibitor cocktail (Roche UK, Burgess Hill, UK)), and sonicated at 4°C using a Bioruptor™ (Diagenode, Liege, Belgium) to generate fragments an average length of 400-500 bp (10 min with 30 s “ON” and “OFF” cycles, power setting high). Kapa hyper prep Kit
attributes.tag: GEO Accession
attributes.value: GSM2974457