Author: Zhou, Jie; Li, Cun; Sachs, Norman; Chiu, Man Chun; Wong, Bosco Ho-Yin; Chu, Hin; Poon, Vincent Kwok-Man; Wang, Dong; Zhao, Xiaoyu; Wen, Lei; Song, Wenjun; Yuan, Shuofeng; Wong, Kenneth Kak-Yuen; Chan, Jasper Fuk-Woo; To, Kelvin Kai-Wang; Chen, Honglin; Clevers, Hans; Yuen, Kwok-Yung
Title: Differentiated human airway organoids to assess infectivity of emerging influenza virus Document date: 2018_6_26
ID: z637eh2z_24
Snippet: Plaque Assay. A plaque assay was performed to determine titers of the virus stocks and supernatant samples as described elsewhere (26) , with minor modifications. Briefly, MDCK cells were seeded in 12-well plates. Confluent monolayers were inoculated with 400 μL of 10-fold serial dilutions of samples and were incubated for 1 h at 37°C. After the inoculum was removed, the monolayers were overlaid with 1% low melting point agarose (Invitrogen) su.....
Document: Plaque Assay. A plaque assay was performed to determine titers of the virus stocks and supernatant samples as described elsewhere (26) , with minor modifications. Briefly, MDCK cells were seeded in 12-well plates. Confluent monolayers were inoculated with 400 μL of 10-fold serial dilutions of samples and were incubated for 1 h at 37°C. After the inoculum was removed, the monolayers were overlaid with 1% low melting point agarose (Invitrogen) supplemented with Eagle's minimal essential medium (MEM) and 1 μg/μL TPCK-treated trypsin and were further incubated for 2-3 d. The monolayers were fixed with 4% paraformaldehyde (PFA) and were stained with 1% crystal violet to visualize the plaque after the agarose plugs were removed. Virus titers were calculated as plaque-forming units per milliliter.
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