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_57
Snippet: The day prior to infection, 4 x 10 5 HeLa cells were plated in 12 well plates with 45 wells per virus 391 (9 time points and 5 replicates per time point). Cells were infected at an MOI of 1 in 150µl 392 volume and incubated on ice for 1 hour with occasional rocking. At one hour, the inocula were 393 aspirated, each well was washed twice with ice cold PBS, and 1ml of fresh, pre-warmed growth 394 media were applied to all wells. One set of 5 wells.....
Document: The day prior to infection, 4 x 10 5 HeLa cells were plated in 12 well plates with 45 wells per virus 391 (9 time points and 5 replicates per time point). Cells were infected at an MOI of 1 in 150µl 392 volume and incubated on ice for 1 hour with occasional rocking. At one hour, the inocula were 393 aspirated, each well was washed twice with ice cold PBS, and 1ml of fresh, pre-warmed growth 394 media were applied to all wells. One set of 5 wells was immediately frozen as the t=0h time 395 point. All other plates were returned to the incubator, and a set of 5 wells was removed and 396 frozen at t=1.5, 2, 2.5, 3, 3.5, 4, 5, and 7 hours. All samples were titered by TCID 50 . The growth 397 curve on PVR-3T3 cells was performed using a similar protocol, except that 5 x 10 5 cells were 398 plated the day prior and the time points were t=1, 2, 3, 4, 5, 6, 7, and 8 hours.
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