GSM5939727: HEK293T_siRNA2; Homo sapiens; OTHER
total RNA were extracted by TRIZOL following the manufacturer's instructions Genomic DNA was digested with DNase I (Ambion), and total RNA was fragmented by heating at 95 °C for 25 mins. The first strand cDNA was transcribed by an anchored oligo d(T) tagged with Illumina A adaptor and T7 promoter, and then the second strand was synthesized to perform in vitro transcription. The second round of reverse transcription was performed with random primers, and then cDNA was used as a template to perform PCR with 15 cycles to amplify the library. Finally, 200-500 bp fragments of the PCR product purified with AMPure XP Beads (Beckman Coulter) were selected for size selection. The final pooled libraries were quantified and sequenced with HiSeq 2500.