title: RNA-seq of embryonic fibroblasts cell lines derived from CC001, CC071 and C57BL/6J mouse strains infected with ZIKV virus
design: RNA-seq of embryonic fibroblasts cell lines derived from CC001, CC071 and C57BL/6J mouse strains infected with ZIKV virus
library_construction_protocol: Embryonic fibroblasts cell lines were infected with ZIKV virus (MOI=5). Total cellular RNA was extracted at 16, 24 and 32 hours post-infection. Mock-infected cells were analyzed 24 hours after mock-infection and used as controls. Three biological replicates (cell lines derived from different embryos) were analyzed for each mouse strain. Cells were lysed in 350 μL RLT buffer (Qiagen) supplemented with 1% β-mercaptoethanol. Total RNA extraction was performed with the RNeasy Mini Kit (Qiagen) according to the manufacturer‟s instructions. On-column digestion with DNase I (Qiagen) was performed to avoid genomic DNA contamination. MEF RNA integrity and quantification were assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). One microgram of high-quality RNA samples (RIN > 9.2) representing biological triplicates were submitted to Novogene for RNA-sequencing (Novogene Beijing, China).  Poly-A selected RNA was used for paired-end library preparation and transcriptome sequencing. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA)