RNA-dependent RNA polymerase speed and fidelity are not the only determinants of the mechanism or efficiency of recombination

Hyejeong Kim; Victor D. Ellis; Andrew Woodman; Yan Zhao; Jamie J. Arnold; Craig E. Cameron

Purified RNA in RNase-free H 2 O was transfected into either HeLa or L929 fibroblasts using 350 TransMessenger (Qiagen). Virus yield was quantified by plaque assay. Briefly, cells and media were 351 harvested at 2-3 days post-transfection, subjected to three freeze-thaw cycles, and clarified. Supernatant 352 was then used on fresh HeLa cells in 6-well plates; virus infection was allowed to continue for 30 min. Luciferase assays. Subgenomic luciferase assays were performed as described previously (32). The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/769224 doi: bioRxiv preprint was isolated 24 h post-infection and used for qRT-PCR to determine genomes/mL or plaque assay to 579 determine pfu/mL, with the quotient yielding specific infectivity, genomes/pfu. (C) Ribavirin 580 sensitivity. HeLa cells were infected at a MOI 0.1 with each PV in the presence of 600 µM ribavirin.