Author: Bouvette, Jonathan; Korkut, Dursun Nizam; Fouillen, Aurélien; Amellah, Soumiya; Nanci, Antonio; Durocher, Yves; Omichinski, James G.; Legault, Pascale
Title: High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension Document date: 2018_12_6
ID: 012ipcdr_36
Snippet: Transfection efficiency was monitored by substituting 5% of the total plasmid transfected by a pTT5SH8Q1-GFPq plasmid [49] . After 48 h, live cells were diluted with one volume of Freestyle 17 media supplemented with 20 mM HEPES pH 7.5, and a thin layer of diluted cells was carefully deposited on a Lab-TEK-II chamber slide. The slide was visualized by both phase contrast microscopy and by fluorescence microscopy on a Nikon TE2000U microscope (20Ã.....
Document: Transfection efficiency was monitored by substituting 5% of the total plasmid transfected by a pTT5SH8Q1-GFPq plasmid [49] . After 48 h, live cells were diluted with one volume of Freestyle 17 media supplemented with 20 mM HEPES pH 7.5, and a thin layer of diluted cells was carefully deposited on a Lab-TEK-II chamber slide. The slide was visualized by both phase contrast microscopy and by fluorescence microscopy on a Nikon TE2000U microscope (20× magnification). The ratio of cells expressing GFP over the total cells of at least 200 cells was taken as the transfection efficiency.
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