Author: Mateo, Roberto; Nagamine, Claude M.; Kirkegaard, Karla
Title: Suppression of Drug Resistance in Dengue Virus Document date: 2015_12_15
ID: 6bx2nrui_38
Snippet: Drug passages. A total of 7 ϫ 10 6 BHK cells on 60-mm tissue culture dishes were pretreated with increasing concentrations of MK-0608 as indicated (or with 0.5% DMSO as a negative control) for 30 min at 37°C and then infected with two independent pools at an MOI of less than 0.1 FFU/cell. Briefly, 0.15 ml of virus in DMEM plus 5% bovine serum was applied to cells, and plates were incubated at 37°C for 1 h. Following infection, medium with or w.....
Document: Drug passages. A total of 7 ϫ 10 6 BHK cells on 60-mm tissue culture dishes were pretreated with increasing concentrations of MK-0608 as indicated (or with 0.5% DMSO as a negative control) for 30 min at 37°C and then infected with two independent pools at an MOI of less than 0.1 FFU/cell. Briefly, 0.15 ml of virus in DMEM plus 5% bovine serum was applied to cells, and plates were incubated at 37°C for 1 h. Following infection, medium with or without MK-0608 was added. After 48 h, supernatants and cells were harvested. Supernatants were used to infect fresh monolayers of BHK cells. This cycle was repeated for 22 passages, using an MOI of less than 0.1 FFU per cell for each passage and increasing concentrations of MK-0608. Pooled viral RNA from each resulting population was sequenced.
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