GSM7158867: Nascent-seq SRSF5 depletion T16 Replicate 2; Homo sapiens; ssRNA-seq
The reaction was stopped by applying TRIzol™RI reagent to the culture dishes. RNA extraction and pulldown of newly transcribed RNA was performed according to (Gressel et al., 2019, PMID: 31399571) without fractionation of the RNA by sonification. Nascent RNA was purified using RNA Clean and Concentrate Kit (ZymoResearch). cDNA libraries for RNA-seq were prepared with universal Plus™ Total RNA-Seq Library Preparation Kit (Tecan) according to manufacturer's instructions. Ribosomal RNA fragments were removed and the library was sequenced on an Illumina NovaSeq6000 instrument strand-specific