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

Article DOI: 10.3791/3702-v • 08:55 min • March 26th, 2012
March 26th, 2012 •
1, 1
1RNA Biology, New England Biolabs

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Summary
Automatic Translation

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 Capping Gaussia Luciferase MRNA HeLa Cell Transfection RNA Polymerase DNA Template Promoter Sequence Transcription Reaction Ribonucleotide Triphosphates RNase Inhibitor Mg2+ Ions RNA Structure And Function Studies Splicing RNAi Experiments Antisense RNA Amplification Microarray Analysis RNA Vaccine Studies Radiolabeled Probes Dye Labeled Probes T7 High Yield RNA Synthesis Kit Linearized Plasmid DNA PCR Products Synthetic DNA Oligonucleotides 5' End Cap Structure
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