GSM8291461: MRC5_KD, no UV, 10 minutes, GRO-seq; Homo sapiens; RNA-Seq
Transcription-competent nuclei were extracted using the Nuclei Isolation Kit. Nuclear Run-On reactions were carried out by adding of Run-On Buffer and incubated for 5 minutes. Br-UTP run-on labeled RNA was isolated using anti-Br-UTP coupled agarose beads. RNA was eluated and purified by phenol-chloroform. The purified RNA was used for the preparation of strand-specific RNA libraries using the TruSeq Stranded Total RNA LT sample Prep Kit, and sequenced under Illumina NovaSeq 6000 system as single-ended 50 bp reads.