title: GSM9050967: A-BOSN12-COSN123_A6_Bru; Mus musculus; OTHER
library_construction_protocol: RNA was extracted using the Quick-RNA Miniprep kit (Zymo) Sequencing libraries were prepared according to Paulsen et al 2014 Methods (PMID: 23973811). Briefly, RNA was was fragmented by heating and then BrU-labeled RNA purified using anti-BrdU antibodies and Protein A-conjugated beads.Nascent RNA was then fragmented at 85C for 10 min and converted to cDNA using NEBNext Ultra II RNA First Strand Synthesis Module (NEB) and NEBNext Ultra II Directional Second Strand Synthesis Kit (NEB) according to manufacturer's instructions. cDNA fragments were then constructed into libraries using NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB).  was then amplified using NEBNext Multiplex Oligos for Illumina (NEB). Libraries were pooled and sequenced on a Novaseq6000 machine.