GSM7034509: MEF IRF3-KO, 4sU, mock, biol rep 4; Mus musculus; RNA-Seq
Samples were lysed in TRIzol and total RNA was extracted using the DirectZOL Microprep kit (Zymo Research). For SLAM conversion, RNA was mixed with 20 mM Iodacetamide (Thermo Fisher Scientific) and incubated at 50°C shaking for 30 min in the dark. The reaction was stopped by mixing with 100 µM dithiothreitol, and converted RNA was purified using the RNA Clean&Concentrator-5 (Zymo Research). 200 ng of converted RNA was used for constructing the libraries. RNA libraries for SLAM-seq were prepared using the TruSeq Stranded mRNA Library Prep kit (Illumina) with oligo-dT beads for capture of poly-A-mRNA following manufacturer's protocols.