title: GSM2859313: JRS4 TEpi MOI 5 infected 6 h rep3; Homo sapiens; RNA-Seq
library_construction_protocol: Medium was aspirated and RNA lysis buffer (Buffer RLT (Qiagen) plus 0.1 M -mercaptoethanol) added to each well. Cells were scraped into the buffer using a cell-scraper. Cell lysate was then loaded onto a QIAshredder homogenizer column (Qiagen, Cat. #79654) and homogenized by centrifugation. RNA was then isolated using the RNeasy Mini Kit (Qiagen, Cat. #74104), according to the manufacturer's instructions. Absorbance of RNA samples was measured at 260 nm using a ND1000 Nanodrop (Biolab), and RNA concentration (µg/ml) was determined by the A260 reading: A260 = 1.0 is equivalent to 40 µg/ml RNA. A260/A280 ratios were measured to determine RNA purity, and an A260/280 ratio of 2.0 – 2.2 was deemed acceptable for subsequent use. Confluent 6-well plates of TEpi cells were infected with JRS4 or 5448 as described above, at an MOI of 5 for 6 h. Mock cells were included as a control. RNA from four biological replicates was extracted as described above. RNA sequencing was performed by The University of Queensland, Institute of Molecular Biosciences (IMB) sequencing facility. Total RNA was deemed as passing quality control tests (2100 bioanalyzer), with a RIN score of >7.0.  For each sample, the mRNA library was prepared using TruSeq stranded mRNA library prep kit (Illumina) and sequenced using 1 x NextSeq 500, 75 cycle (1 x 75bp) Illumina High Output single-end run.
attributes.tag: GEO Accession
attributes.value: GSM2859313