labeling: Nascent transcripts were labeled by incubation with 0.25mM EU for 2 hours following release from nocodazole. 10^6 cells were collected by certrifugation and lysed by addition of Trizol. rRNA was depleted from 5ug of total RNA using the Ribozero kit. EU-labeled RNA was biotinylated and recovered using the Click-IT EU-labeled RNA was converted to directional sequencing libraries using NEBNext Ultra Directional RNA Library Prep Kit. NEBNext Ultra Directional RNA Library Prep Kit
GEO Accession
GSM4147295