title: GSM6924716: HSV-110_Day187; Homo sapiens; RNA-Seq
library_construction_protocol: PBMCs were isolated with Ficoll-Hypaque reagent and lysed in QIAzol (Qiagen, Hilden, Germany) and stored at -80°C until use. Their total RNA was extracted and purified with the miRNeasy kit (Qiagen). RNA quality and quantity were estimated using Nanodrop (Thermo Scientific, Wilmington, DE) and Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). Stranded cDNA sequencing libraries were generated with TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA) following the manufacturer's instructions. Briefly 500 ng of total RNA was used for mRNA selection. After the reverse transcription to 1st strand cDNA, strand info was reserved with dUTPs during 2nd strand synthesis. The dsDNA fragments then had the addition of a single 'A' base and subsequent ligation of the adapter. The products were then purified and enriched with PCR to create the final cDNA library. The library was qualified with Agilent Bioanalyzer and quantified with Qubit 2.0 fluorometer. The cluster generation and pair-end (2x75 bp) sequencing was run on Illumina HiSeq 3000 at NHLBI Sequencing Core. Up to 96 barcoded samples were pooled for one single run, which yielded at least 30M passed filter paired reads per sample.