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JoVE Journal Biochemistry
An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
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An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

Article DOI: 10.3791/61648-v • 09:52 min • September 15th, 2020
September 15th, 2020 •
*1, *1, 1
1Molecular Biology Program, Memorial Sloan Kettering Cancer Center
* These authors contributed equally

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Summary
Automatic Translation

Tracking individual translation events allows for high-resolution kinetic studies of cap-dependent translation mechanisms. Here we demonstrate an in vitro single-molecule assay based on imaging interactions between fluorescently labeled antibodies and epitope-tagged nascent peptides. This method enables single-molecule characterization of initiation and peptide elongation kinetics during active in vitro cap-dependent translation.

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In Vitro Single-Molecule Imaging Assay Cap-Dependent Translation Kinetics Cell-free Translation Systems Bulk Translation Properties Single Molecule Resolution Bulk Translation Assays Cap-dependent Translation Microscope Incubator Laser Power Button Laser Safety Key Imaging Mode Objective Filter Set EMCCD Camera Stage Controller Syringe Pump Computer Software
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