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- Run-off_transcription abstract "A run-off transcription assay is an assay in molecular biology which is conducted in vitro to identify the position of the transcription start site (+1) of a specific promoter along with its accuracy and rate of in vitro transcription. Run-off transcription can be used to quantitatively measure the effect of changing promoter regions on in vitro transcription levels, Because of its in vitro nature, however, this assay cannot accurately predict cell-specific gene transcription rates, unlike in vivo assays such as nuclear run-on.To perform a run-off transcription assay, a gene of interest, including the promoter, is cloned into a plasmid The plasmid is digested at a known restriction enzyme cut site downstream from the transcription start site such that the expected mRNA run-off product would be easily separated by gel electrophoresis.DNA needs to be highly purified prior to running this assay. To initiate transcription, radiolabeled UTP, the other nucleotides, and RNA polymerase are added to the linearized DNA. Transcription continues until the RNA polymerase reaches the end of the DNA where it simply “runs off” the DNA template, resulting in an mRNA fragment of a defined length. This fragment can then be separated by gel electrophoresis, alongside size standards, and autoradiographed. The corresponding size of the band will represent the size of the mRNA from the restriction enzyme cut site to the transcription start site (+1). The intensity of the band will indicate the amount of product mRNA produced.".
- Run-off_transcription wikiPageID "42611813".
- Run-off_transcription wikiPageLength "4499".
- Run-off_transcription wikiPageOutDegree "12".
- Run-off_transcription wikiPageRevisionID "678244285".
- Run-off_transcription wikiPageWikiLink Category:Molecular_biology_techniques.
- Run-off_transcription wikiPageWikiLink Gel_electrophoresis.
- Run-off_transcription wikiPageWikiLink Isotopic_labeling.
- Run-off_transcription wikiPageWikiLink MRNA.
- Run-off_transcription wikiPageWikiLink Messenger_RNA.
- Run-off_transcription wikiPageWikiLink Molecular_biology.
- Run-off_transcription wikiPageWikiLink Nucleotide.
- Run-off_transcription wikiPageWikiLink Plasmid.
- Run-off_transcription wikiPageWikiLink Promoter_(genetics).
- Run-off_transcription wikiPageWikiLink RNA_polymerase.
- Run-off_transcription wikiPageWikiLink Restriction_enzyme.
- Run-off_transcription wikiPageWikiLink Transcription_(genetics).
- Run-off_transcription wikiPageWikiLink Uridine_triphosphate.
- Run-off_transcription wikiPageWikiLinkText "Run-off transcription".
- Run-off_transcription hasPhotoCollection Run-off_transcription.
- Run-off_transcription wikiPageUsesTemplate Template:Multiple_issues.
- Run-off_transcription wikiPageUsesTemplate Template:Reflist.
- Run-off_transcription subject Category:Molecular_biology_techniques.
- Run-off_transcription hypernym Assay.
- Run-off_transcription comment "A run-off transcription assay is an assay in molecular biology which is conducted in vitro to identify the position of the transcription start site (+1) of a specific promoter along with its accuracy and rate of in vitro transcription.".
- Run-off_transcription label "Run-off transcription".
- Run-off_transcription sameAs m.010gpf12.
- Run-off_transcription wasDerivedFrom Run-off_transcription?oldid=678244285.
- Run-off_transcription isPrimaryTopicOf Run-off_transcription.