Total RNA-Seq MHV68 Infection Models

MHV68 de novo infection: NIH 3T3 cells were infected with MHV68 H2B-YFP (wildtype) or ORF50stop at an MOI of 5 PFU per cell. RNA was collected from infected samples at 18 hours post infection. If indicated, cells were treated with 100 ug/mL Phosphonacetic Acid at 1.5 hr post induction. MHV68 lytic reactivation: A20-HE-RIT cells were induced in 1x RPMI media containing 5 ug/mL Doxycycline and 20 ng/mL 12-O-Tetradecanoyl-phorbol-13-acetate (TPA) for 24 hours. Total RNA was isolated from cells using the Direct-zol RNA MiniPrep Kit. ERCC spike-in controls were added. RNA was ribominus selected and directional cDNA libraries were generated using either Stranded Total RNA Prep with Ribo-Zero Plus (Illumina # 20040525) or TruSeq Stranded Total RNA Ribo-Zero Gold (Illumina #RS-122-2303). 2 biological replicates were sequenced for all samples. Sequencing was performed at the NCI CCR Frederick Sequencing Facility using the Illumina NovaSeq SP platform to generate 150 bp PE reads.