title: GSM5259352: A549 cells, LCMV infected at an MOI of 0.01, biological replicate 1; Homo sapiens; RNA-Seq
library_construction_protocol: A549 cells were infected with LCMV at an MOI of 0.01 or mock-treated. At 36 hours post infection, cells were collected and total RNA was extracted with Trizol (Tiangen, Beijing) and RNA level was assessed with Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA, USA) and NanoPhotometer spectrophotometer (Implen). Triplicate samples of all assays were used to construct independent libraries, followed by sequencing and analysis. The NEB Next Ultra RNA Library Prep Kit for Illumina (NEB) was used to construct the libraries for sequencing. NEB Next Poly (A) mRNA Magnetic Isolation Module kit was used to enrich the poly (A) tailed mRNA. The mRNA was fragmented into ~200 base pair pieces. The first-strand cDNA was synthesized from the mRNA fragments using reverse transcriptase and random hexamer primers, and then the second-strand cDNA was synthesized using DNA polymerase I and RNaseH. The end of the cDNA fragment was subjected to an end repair process that included the addition of a single ''A'' base, followed by ligation of the adapters. Products were purified and enriched by polymerase chain reaction (PCR) to amplify the library DNA. The final libraries were quantified using Qubit 2.0 Fluorometer (Invitrogen) and an Agilent 2100 Bioanalyzer. After quantitative reverse transcription-PCR validation, libraries were subjected to paired-end sequencing with pair end 150-base pair reading length on an Illumina Novaseq 6000 (Illumina).