GSM8773917: LIN54-WT, biol rep 1, Stag2-WT mouse urothelium, 1 week after TMX induction; Mus musculus; OTHER
Urothelial cells (approx. 500,000 cells for each experimental condition) were used for CUT&RUN experiments (2-3 replicates per experimental condition). Cut&Run was performed as described by Skene et al. 2018. DNA samples were processed through subsequent enzymatic treatments for end-repair, dA-tailing, and ligation to adapters with "NEBNext Ultra II DNA Library Prep Kit for Illumina" (NEB ref.E7645). Adapter-ligated libraries were completed by a limited-cycle PCR, with 63º T(ann/ext) for 10s, and extracted with a [single] double-sided SPRI size selection. Resulting average fragment size is 320 bp from which 120 bp correspond to adaptor sequences. Libraries were applied to an Illumina flow cell for cluster generation and sequenced on an Illumina instrument by following manufacturer's recommendations.