title: GSM9058801: BasalCells, E16.5, TKO, GFP, Rep1; Mus musculus; OTHER
library_name: GSM9058801
library_construction_protocol: Dorsal skin tissue was dissected from embryos and flash-frozen in liquid nitrogen, which was then followed by dry-pulverization using a Cell Crusher. Tissue powder was then resuspended in NUN buffer. Drosophila tissue was added as spike-in. Chromatin was then fragmented and solubalized with DNase I and nascent RNA was biotin-labeled via run-on reaction. Cell-type-specific nascent RNAs were enriched sequentially through GFP pull-down and streptavidin beads, and then extracted using standard TRIzol extraction protocol. Input samples were aliquoted before GFP pull-down to serve as control. Nascent RNA was then processed for 3'-adaptor ligation, 5' decapping, end-repair, 5'-adaptor ligation, PCR enrichement and finally AMPure XP beads clean-up.