GSM4704573: IFN siCTRL RIBO rep4; Homo sapiens; OTHER
For RNA-seq, cells were harvested in TRIzol and RNA was extracted by manufacturer protocol, after which TURBO DNase treatment was performed and RNA was phenol-chloform precipitated. For Ribo-seq, cells were lysed in polysome lysis buffer and cytoplasmic lysates were clarified by centrifugation, then ribosome protected fragments were generated by micrococcal nuclease treatment. Monosome fractions were isolated by sucrose gradient ultracentrifugation, and RPF RNA was size-selected on TBE urea gels, eluted, T4 PNK treated, and phenol-chloform extracted. For RNA-seq, libraries were prepared using the KAPA Stranded mRNA-Seq Kit. For Ribo-seq, libraries were prepared using the NEB Next Small RNA Library Prep Kit.