GSM9230301: 24132XR-06-04; Homo sapiens; RNA-Seq
Total RNA was extracted using miRNeasy QIAGEN mini kit according to the manufacturer's protocol (QIAGEN #1038703). Totally, 1000 ng of RNA were reverse transcribed using superscript IV Uniprime (Invitrogen #12596025) with random hexamers following the manufacturer's recommendations. cDNA was diluted with RNase and DNase free water prior to use in a quantitative real-time polymerase chain reaction (qRT-PCR). RNA was isolated from samples using RNeasy® Plus Mini Kit (Qiagen, Hilden, Germany) per the manufacturer's instructions. Isolated RNA sample quality was assessed by RNA Tapestation (Agilent Technologies Inc., California, USA) and quantified by Qubit 2.0 RNA HS assay (ThermoFisher, Massachusetts, USA). Libraries were constructed with KAPA™ RNA HyperPrep with RiboErase (Roche, Indiana, USA) and performed based on manufacturer's recommendations. Final library quantity was measured by KAPA SYBR® FAST qPCR and library quality evaluated by TapeStation HSD1000 ScreenTape (Agilent Technologies, CA, USA). The final library size was about 400bp with an insert size of about 250bp. Illumina® 8-nt unique dual-indices were used. Equimolar pooling of libraries was performed based on QC values and sequenced on an Illumina® NovaSeq (Illumina, California, USA) with a read length configuration of 150 PE for 60M PE reads per sample (30M in each direction).