This content is Free Access.
בתעתיק במבחנה תקרת של mRNA בלוציפראז Gaussia ואחריו transfection Cell הלה
17,864 Views
•
08:55 min
•
March 26, 2012
DOI:
Summary
Automatically generated
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.
Explore More Videos
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
Read Article
Cite this Article
Jani, B., Fuchs, R. In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection. J. Vis. Exp. (61), e3702, doi:10.3791/3702 (2012).
Copy