When the cell density in the cell culture flask reached 70–80%, the H226 cells were digested with trypsin. Cells were collected by centrifugation at 40 × g, and then the cells were placed in the 24-well plate to make the density reach ~60% after resuspended. After mixture, the cells continued to be cultured in an incubator at 37°C. Plasmid transfection was performed using the TurboFect™ transfection kit (Fermentas, Inc.), and fresh medium was replaced 12 h after transfection. After 72 h of transfection, the cells were washed twice with PBS, and 250 µl of 1X PLB lysis buffer (Thermo Fisher Scientific, Inc.) was then added to each well. LAR II reagent (100 µl) and lysate (20 µl) were added into a 96-well plate, and then 100 µl of Stop & Glo substrate (Promega Corp.) was added within 10 sec to measure the luciferase activity. After exporting the data, the results were analyzed and plotted using GraphPad Prism 5 software (GraphPad Software, Inc.).
Turbofect transfection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
TurboFect™ transfection kit is a laboratory reagent designed for the transfection of nucleic acids, such as plasmid DNA, siRNA, or mRNA, into a variety of cell types. It is a cationic polymer-based transfection agent that forms complexes with nucleic acids, facilitating their uptake and expression in the target cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using turbofect transfection kit
1
Plasmid Transfection and Luciferase Assay
2
DENV2 Infection of Transfected Huh7 Cells
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Huh7 cells (3 × 105 cells/ well) were plated into the 6-well plate, and the transfection was performed following the manufacturer’s instructions of Turbofect transfection kit (Fermentas, Carlsbad, CA, USA). The transfected Huh7 cells were infected with DENV2 at a MOI of 10 at 18 h post transfection at 37 °C for 2 h and replaced with 10% FBS DMEM. The samples were collected at 36 h p.i. for different experiments.
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