GSM7991570: T24, siYY1, biol rep 2 (Y2); Homo sapiens; RNA-Seq
Total RNAs were extracted from samples using TRIzol (Invitrogen). DNA digestion was carried out after RNA extraction by DNaseI. RNA quality was determined by examining A260/A280 with Nanodrop OneCspectrophotometer (Thermo Fisher Scientific Inc). RNA Integrity was confirmed by 1.5% agarose gel electrophoresis. Qualified RNAs were finally quantified by Qubit3.0 with Qubit RNA Broad Range Assay kit (Life Technologies). 2 μg total RNAs were used for stranded RNA sequencing library preparation using KC-Digital Stranded mRNA Library Prep Kit for Illumina (Catalog NO. DR08502, Seqhealth Technology Co., Ltd. Wuhan, China) following the manufacturer's instruction. The kit eliminates duplication bias in PCR and sequencing steps, by using unique molecular identifier (UMI) of 8 random bases to label the pre-amplified cDNA molecules.