title: GSM6119036: Total RNA-seq Patient 2T; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from samples using TRIzol reagent (Invitrogen) according to the manufacturers instructions. For Chipseq, DNA samples were purified using UPrep® Spin Columns (Genesee Scientific). Total RNA-seq (ribo-depleted) libraries for all samples were generated and sequenced by Admera Health. Briefly, isolated RNA sample quality was assessed by High Sensitivity 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 manufacturers recommendations. Final library quantity was measured by KAPA SYBR® FAST qPCR and library quality evaluated by TapeStation D1000 ScreenTape (Agilent Technologies, CA, USA). The final library size was about 430 bp with an insert size of about 200bp. Illumina® 8-nt dual-indices were used. Equimolar pooling of libraries was performed based on QC values and sequenced on an Illumina® NovaSeq S4 (Illumina, California, USA) with a read length configuration of 150 PE for 50 M PE reads per sample (25M in each direction).