title: In vitro assay of pre-mRNA splicing in mammalian nuclear extract.
abstract: The in vitro splicing assay is a valuable technique that can be used to study the mechanism and machinery involved in the splicing process. The ability to investigate various aspects of splicing and alternative splicing appears to be endless due to the flexibility of this assay. Here, we describe the tools and techniques necessary to carry out an in vitro splicing assay. Through the use of radiolabeled pre-mRNA and crude nuclear extract, spliced mRNAs can be purified and visualized by autoradiography for downstream analysis.
authors.affiliation: Department of Microbiology & Molecular Genetics, University of California, Irvine, CA, USA.
authors.forename: Maliheh
authors.initials: M
authors.lastname: Movassat
authors.forename: William F
authors.initials: WF
authors.lastname: Mueller
authors.forename: Klemens J
authors.initials: KJ
authors.lastname: Hertel
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D002457:Cell Extracts / Q000235:genetics / Q000302:isolation & purification; D002467:Cell Nucleus / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008322:Mammals; D008967:Molecular Biology / Q000379:methods*; D012322:RNA Precursors / Q000235:genetics*
chemical_list: D002457:Cell Extracts; D012322:RNA Precursors