GSM5592459: 2B4zIsecrIL15_donor2; Homo sapiens; RNA-Seq
RNA was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany; cat. 74104). RNA preparation included an on-column RNase-Free DNase digestion (Qiagen, Hilden, Germany;cat. 79254). RNA samples were converted to double stranded cDNA using the Ovation RNA-Seq System v2.0 kit (Tecan Cat. No. / ID: 7102-32), which utilizes a proprietary strand displacement technology for linear amplification of mRNA without rRNA/tRNA depletion as per the manufacturer's recommendations.  This approach does not retain strand specific information. Quality and quantity of the resulting cDNA was monitored using the Bioanalyzer High Sensitivity kit (Agilent Cat. No. / ID: 5067-4626) which yielded a characteristic smear of cDNA molecules ranging in size from 500 to 2000 nucleotides in length.  After shearing 500 nanograms of cDNA to an average size of 250 nucleotides with the Covaris S4 (Covaris Inc. Woburn, MA) library construction was completed with the Illumina Truseq Nano kit (Cat. No. / ID:   20015964) according to the manufacturer's instructions. mRNA libraries were sequenced on an Illumina Novaseq 6000 instrument using 150bp paired-end dual indexed reads and 1% of PhiX control.