title: GSM1480812: subject 169, ADEH-, HSV-1 treatment, rep 1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated from PBMCs using RNeasy Mini Kits (Qiagen, Valencia, CA) according to the manufacturer’s guidelines. RNA concentration and purity were determined using Nano Drop-1000 Spectrophotometer and Agilent 2100 Bioanalyzer RNA Nano Chip (Agilent Technol., Palo Alto, CA).  Library preparation and HiSeq sequencing were performed at The University of Colorado Denver Genomics and Microarray Core facility. A total of 200-1000 ng of total RNA was used to prepare the Illumina HiSeq libraries according to manufacturer’s instructions for the TruSeq RNA kit.  In brief, the poly-A containing mRNA molecules were purified using poly-T oligo-attached magnetic beads.  Following purification, the mRNA was fragmented into small pieces using divalent cations under elevated temperature.  The cleaved RNA fragments were converted into first strand cDNA using reverse transcriptase and random primers.  This was followed by second strand cDNA synthesis using DNA polymerase I and RNase H. These cDNA fragments then went through an end repair process, the addition of a single “A” base, and then ligation of the adapters. RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM1480812