title: GSM2194159: RYBPfl/fl.YAF2KO_nucRNA_OHT_rep3; Mus musculus; RNA-Seq
library_construction_protocol: 1 x 10^7 Rybpfl/fl;Yaf2-/- cells with or without tamoxifen were resuspended in 1 mL HS Lysis buffer (50 mM KCl, 10 mM MgSO4.7H20, 5 mM HEPES, 0.05% NP40 (IGEPAL CA630), 1 mM PMSF, 3 mM DTT) for 1 min at room temperature to isolate nuclei. Nuclei were centrifuged at 1,000 x g for 5 min at 4°C, followed by a total of three washes with ice-cold RSB buffer. Nuclei were then resuspended in 1mL of TriZOL reagent (Thermo Scientific) according to the manufacturer's protocol. Nuclear RNA was treated with the TURBO DNA-free Kit (Thermo Scientific) and depleted for rRNA using the RiboMinus kit and protocol (Thermo Scientific). RNA-seq libraries were prepared using the NEBNext Ultra Directional RNA-seq kit (NEB) and libraries were sequenced on the Illumina NextSeq 500 with 40bp paired-end reads.
attributes.tag: GEO Accession
attributes.value: GSM2194159