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Capitoli
- 00:05Titolo
- 01:33In vitro Transcription using the T7 High-yield RNA Synthesis Kit
- 03:29RNA Capping using the Vaccinia Capping System
- 04:17HeLa Cell Transfection using the TransIT mRNA Transfection Kit
- 06:08Gaussia Luciferase (GLuc) and Cypridina Luciferase (CLuc) Assays
- 07:20Representative Results
- 08:36Conclusion
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 TranscriptionCappingGaussia Luciferase MRNAHeLa Cell TransfectionRNA PolymeraseDNA TemplatePromoter SequenceTranscription ReactionRibonucleotide TriphosphatesRNase InhibitorMg2+ IonsRNA Structure And Function StudiesSplicingRNAi ExperimentsAntisense RNA AmplificationMicroarray AnalysisRNA Vaccine StudiesRadiolabeled ProbesDye Labeled ProbesT7 High Yield RNA Synthesis KitLinearized Plasmid DNAPCR ProductsSynthetic DNA Oligonucleotides5′ End Cap Structure
