GSM2667361: Patient 24, conv sample, technical replicate 2; Homo sapiens; RNA-Seq
Whole blood collected in PAXgene Boold RNA Tube Total RNA was extracted from PAXgene RNA blood with the PAXgene Blood RNA Kit (Qiagen) by following manufacturers’ instructions including DNase digestion and an additional clean-up using RNeasy MinElute kit (Qiagen). Purified RNA samples were quantified by Qubit 3.0 fluorometer with RNA High Sensitivity Assay kit (Thermo-Fisher). The quality of the RNA was confirmed by TapeStation 2200 with the RNA High Sensitivity ScreenTape (Agilent Technologies). To prepare the 96 samples’ libraries, ribosomal RNA (rRNA) and globin mRNA were removed from 200ng total RNA, and the remaining RNA was fragmented and primed for cDNA synthesis using TruSeq Total Stranded RNA HT kit with Ribo-Zero Globin on a Microlab STAR automated liquid handling system (Hamilton). The libraries were barcoded with TruSeq HT indices to allow for multiplexing and ligation-mediated PCR was performed to enrich barcoded libraries for 15 cycles, then purified with the Agencourt AMPure XP beads system (Beckman Coulter). The libraries were assessed for quality with the high sensitivity DNA chip in a TapeStation 2200 (Agilent) and quantified with KAPA Library Quantification Kits for Illumina platforms (Kapa Biosystems). The libraries were diluted to 2nM and combined equimolarly in pools of 12.
GEO
GSM2667361
GEO Accession
GSM2667361