Selected article for: "cell assay and cytopathic effect"
Author: Laura Riva; Shuofeng Yuan; Xin Yin; Laura Martin-Sancho; Naoko Matsunaga; Sebastian Burgstaller-Muehlbacher; Lars Pache; Paul P. De Jesus; Mitchell V. Hull; Max Chang; Jasper Fuk-Woo Chan; Jianli Cao; Vincent Kwok-Man Poon; Kristina Herbert; Tu-Trinh Nguyen; Yuan Pu; Courtney Nguyen; Andrey Rubanov; Luis Martinez-Sobrido; Wen-Chun Liu; Lisa Miorin; Kris M. White; Jeffrey R. Johnson; Christopher Benner; Ren Sun; Peter G. Schultz; Andrew Su; Adolfo Garcia-Sastre; Arnab K. Chatterjee; Kwok-Yung Yuen; Sumit K. Chanda
Title: A Large-scale Drug Repositioning Survey for SARS-CoV-2 Antivirals
  • Document date: 2020_4_17
    • ID: 1fgnfh62_15
    Snippet: Compounds from the LOPAC®1280 and ReFRAME library were transferred into F-BOTTOM, µCLEAR®, BLACK 384-well plates (Greiner) using an Echo 550 Liquid Handler (Labcyte). All compounds were diluted in culture media to a final concentration of 5 µM during screening. Briefly, Vero E6 cells were seeded in 384-well plates, on top of pre-spotted compounds, at a density of 3,000 cells per well in 40 µl using a microFlo™ select dispenser (BioTek Inst.....
    Document: Compounds from the LOPAC®1280 and ReFRAME library were transferred into F-BOTTOM, µCLEAR®, BLACK 384-well plates (Greiner) using an Echo 550 Liquid Handler (Labcyte). All compounds were diluted in culture media to a final concentration of 5 µM during screening. Briefly, Vero E6 cells were seeded in 384-well plates, on top of pre-spotted compounds, at a density of 3,000 cells per well in 40 µl using a microFlo™ select dispenser (BioTek Instruments). Sixteen hours post-seeding, the cells were infected by adding 10 µl of SARS-CoV-2 per well at an MOI of 0.01. Cytopathic effect (CPE) was indirectly quantified as the presence of ATP in live cells by using the CellTiter-Glo (Promega) luminescent cell viability assay at 72 hours post-infection. Data were normalized to the median of each plate. For the ReFRAME library, the Z-score was calculated based on the log2FC with the average and standard deviation of each plate. The screen was performed in duplicate by running the assay in parallel for the LOPAC®1280 library or as two independent experiments for the ReFRAME collection.

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