RNA-Seq of Murine Fibroblasts  (Mock Infected) (1 hpi)
Total RNA was isolated from  infected L929 cells at 1 hour and 3 hours post infection (hpi) using the  TRIZol Plus RNA Isolation Kit (Life Technologies) according to the  manufacturers directions. To ensure that the highest quality RNA is used in  this analysis, 4 biological replicates of each condition were isolated. RNA  was quantitated via nanodrop and bioanalyzer. Libraries were prepared using  the Illumina TruSeq strand specific mRNA sample preparation system  (Illumina).Briefly, mRNA was extracted from total RNA using polyA selection,  followed by RNA fragmentation. Strand specific library was constructed by  first-strand cDNA synthesis using random primers, sample cleanup and  second-strand synthesis using DNA Polymerase I and RNase H. A single 'A' base  was added to the cDNA fragments followed by ligation of the adapters. Final  cDNA library was achieved by further purification and enrichment with PCR,  quality checked using Agilent 4200 TapeStation. The libraries were sequenced  (Single end 100bp reads) using the Illumina HiSeq4000, with 10 samples per  lane, and final of over 20 million reads per sample.