Author: William Fitzsimmons; Robert J. Woods; John T. McCrone; Andrew Woodman; Jamie J. Arnold; Madhumita Yennawar; Richard Evans; Craig E. Cameron; Adam S. Lauring
Title: A speed-fidelity trade-off determines the mutation rate and virulence of an RNA virus Document date: 2018_4_27
ID: 8p24gszj_41
Snippet: In vitro transcription, transfection, and viral stocks 329 Viral RNA was generated by in vitro transcription of the corresponding plasmid clone using T7 330 RNA polymerase, and virus was recovered following RNA transfection of HeLa. For 331 transfections, 2.6 x 10 5 HeLa were plated per well in a 12 well plate the day prior. One The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101.....
Document: In vitro transcription, transfection, and viral stocks 329 Viral RNA was generated by in vitro transcription of the corresponding plasmid clone using T7 330 RNA polymerase, and virus was recovered following RNA transfection of HeLa. For 331 transfections, 2.6 x 10 5 HeLa were plated per well in a 12 well plate the day prior. One The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 14 0.01. For all stocks, cells were subjected to three freeze-thaw cycles and the supernatants 338 clarified by centrifugation a 1400 x g for 4 minutes. These supernatants were stored at minus 339 80ËšC in aliquots to limit the number of subsequent freeze-thaw cycles.
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