title: Illumina NovaSeq 6000 paired end sequencing; Transcriptome analysis of a novel alveolar porcine epithelial cells line infected with different live attenuated Influenza Viruses
design: Transcriptome analysis of a novel alveolar porcine epithelial cells line infected with different live attenuated Influenza Viruses
library_construction_protocol: At 24 or 48 hours p.i., the cells were washed twice and cells were lysed using TRIzol Reagent (Life Technologies). The T3 porcine bronchiolar epithelial cell line was generated by InSCREENeX GmbH (Braunschweig, Germany) and kindly provided by Sasha Trapp (Institut national de la recherche agronomique, INRA, France). The cells were maintained at 37°C and 5% CO2 in porAEC medium with supplement (InSCREENeX, INS-ME-1024) in collagen-coated (InSCREENeX, INS-SU-1017) flasks Cells grown in 24-well cell culture plates were inoculated with virus using an MOI of 1 TCID50/cell. Poly I:C was used at a concentration 1/10000. Following an incubation for 90 minutes at 37°C, the inoculum was removed, the cells washed three times with PBS, and fresh porAEC medium added, then cells were returned to the CO2 incubator at 37°C. RNA was extracted using NucleoSpin RNA  (Macherey Nagel cat# 740955.50). Library preparation was performed using BRB-Seq library preparation kit (Alithea genomics, PN 10813) following manufacturer's instruction.