In Vitro Splicing Assays

Akila Mayeda, Adrian R. Krainer

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

Abstract

In vitro splicing assays represent an important technique to analyze, in detail, the basic splicing mechanisms for a pre-mRNA of interest. The cell-free assays, under defined conditions, have many advantages for characterizing the cis-acting elements and trans-acting factors responsible for the regulation of alternative splicing. Radiolabeled transcripts of minigenes of interest are spliced in HeLa cell nuclear extracts under various conditions, and the splicing products analyzed directly using denaturing polyacrylamide gel electrophoresis, or detected indirectly using RT-PCR. Here, a standard method of in vitro splicing is described that has been used successfully for many years in the authors' laboratories.

Original languageEnglish
Title of host publicationAlternative pre-mRNA Splicing
Subtitle of host publicationTheory and Protocols
PublisherWiley-VCH
Pages320-329
Number of pages10
ISBN (Print)9783527326068
DOIs
Publication statusPublished - 02-02-2012

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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    Mayeda, A., & Krainer, A. R. (2012). In Vitro Splicing Assays. In Alternative pre-mRNA Splicing: Theory and Protocols (pp. 320-329). Wiley-VCH. https://doi.org/10.1002/9783527636778.ch30