GSM5710634: pre-gRNA_CDR1as_KD-rep2; Homo sapiens; RNA-Seq
RNA extraction was started by lysing the cells in TRIzol reagent (Invitrogen) and followed by the usage of Direct-zol RNA Miniprep Kit (ZYMO RESEARCH, CA, USA) according to the manufacturer's instructions including DNase on-column treatment for 15 min at room temperature. RNA concentrations and purity were determined using Nanodrop Lite (Thermo Fisher Scientific). RNA concentrations were measured using the Qubit RNA HS Assay Kit (Thermo Fisher  Scientific) on a Qubit 4 Fluorometer (Thermo Fisher Scientific), as well as using Nanodrop Lite (Thermo Fisher Scientific). RNA quality was checked using Agilent Bioanalyzer 2100 (Agilent Technologies) and all RNA integrity (RIN) values exceeded 8.0. Library preparation, and poly(A) RNA sequencing was performed by Beijing Genomics Institute (BGI) in Copenhagen. Briefly, 1 mg total RNA input was used. Non-stranded and polyA-selected RNA library preparation was applied and PE100 sequencing was performed following the specific protocol for sequencing on a DNBSEQ sequencing platform (MGI, Shenzhen, China). Library size and purity control was performed on a Agilent 2100 Bioanalyzer (Agilent Technologies) using a High-Sensitivity DNA-Chip.