GSM9824067: OVCAR-5-CisR siNUDT5 replicate 1; Homo sapiens; RNA-Seq
RNA was extracted using TRIzol reagent (Thermo Fisher Scientific, 15596026) and the RNeasy Mini Kit (Qiagen, 74104) according to the manufacturer's instructions. RNA quality and concentration were assessed by spectrophotometry (NanoDrop 8000, Thermo Fisher Scientific). Libraries for RNA-Seq were prepared with KAPA Stranded RNA-Seq Kit with RiboErase Kit. Ribosomal RNA was depleted by hybridization of complementary DNA oligonucleotides, followed by treatment with RNase H and DNase and RNA fragmentation. First-strand cDNA synthesis was performed using random priming, followed by second-strand synthesis to convert cDNA:RNA hybrid to double-stranded cDNA and incorporates dUTP into the second cDNA strand. cDNA generation is followed by A-tailing, adaptor ligation, and PCR amplification.