title: GSM4116224: DM2-LR_PBMC_1161; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA from DNA/RNA Shield Blood Collection Tubes was isolated by using Quick-DNA/RNA Blood Tube Kit with DNase treatment according to manufacturer's provided protocol (Zymo Research). Total RNA from PBMCs was isolated by using TRIzol Reagent and the Direct-zol RNA MiniPrep Kit with DNase treatment according to manufacturer's provided protocol (Zymo Research). Total RNA from mouse thymus was isolated using PolyTron (Kinematica) homogenization in TRI Reagent (Sigma) followed by treatment with the RNeasy Kit (Qiagen) according to the manufacturer's instructions. RNA was quantitated on a Nanodrop, Quant-iT RiboGreen RNA Assay Kit (Thermo Fisher) and/or Qubit RNA BR Assay Kit (Thermo Fisher Scientific). Thymic RNA-seq (2 × 50 paired-end reads) were performed by The New York Genome Center from total RNA. For human PBMC and whole blood strand-specific, rRNA- and globin- (only whole blood) depleted RNA-seq libraries were prepared using the KAPA Stranded RNA-seq Kit with RiboErase HMR Globin (Kapa Biosystems) per manufacturer's instructions, except for the use of custom Illumina-compatible index primers to allow multiplexing. Library size distribution was assessed using the High Sensitivity NGS Fragment Analysis Kit (DNF-747) on a Fragment Analyzer (Agilent) and 2 × 76 paired-end sequencing was performed using an Illumina NextSeq 500.
attributes.tag: GEO Accession
attributes.value: GSM4116224