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Trascrizione in vitro e tappatura mRNA Luciferase Gaussia Seguito da cellule HeLa Transfection
<em>In vitro</em> Transcription and Capping of <em>Gaussia</em> Luciferase mRNA Followed by HeLa Cell Transfection
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection

Trascrizione in vitro e tappatura mRNA Luciferase Gaussia Seguito da cellule HeLa Transfection

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08:55 min

March 26, 2012

DOI:

08:55 min
March 26, 2012

18209 Views
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Summary

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This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene. The RNA is purified and a fraction of the uncapped RNA is enzymatically capped using the Vaccinia virus capping enzyme. In the final step, the mRNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity.

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