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In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection

DOI :

10.3791/3702-v

8:55 min

March 26th, 2012

March 26th, 2012

17,711 Views

1RNA Biology, New England Biolabs

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.

Tags

In Vitro Transcription

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