title: GSM3731243: mES_0.1Gy_8h_1; Mus musculus; OTHER
library_construction_protocol: For Bru-seq experiments, we followed the previously published protocol from Paulsen et al., Methods, 2014. Briefly, bromouridine (BrU) was added to the culture media at a final concentration of 2mM and the dishes were returned to CO2 incubator at 37°C for 30 minutes to label the nascent RNA. Labeling was done in the last 30 minutes of the corresponding incubation times for each experiment. Cells were trypsinized, collected pellets were frozen in liquid nitrogen and stored at -80°C until use. From then on BrU-seq protocol was applied. Cell pellets were then lysed in Trizol, total RNA isolated, BrU-RNA was immunoprecipitated using anti-BrdU antibodies (BD Pharmingen, 555627), strand-specific DNA libraries were prepared with Illumina TruSeq kit [Illumina, RS-122-2001]. Samples were then deep sequenced using Illumina sequencing platform. Library construction was performed according to the previously published protocol from Paulsen et al., Methods, 2014. BrU-seq
attributes.tag: GEO Accession
attributes.value: GSM3731243