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

Fingerprint

Assays
Trans-Activators
RNA Precursors
Alternative Splicing
Cell Extracts
Electrophoresis
HeLa Cells
Polyacrylamide Gel Electrophoresis
Polymerase Chain Reaction
In Vitro Techniques
polyacrylamide gels

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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
Mayeda, Akila ; Krainer, Adrian R. / In Vitro Splicing Assays. Alternative pre-mRNA Splicing: Theory and Protocols. Wiley-VCH, 2012. pp. 320-329
@inbook{aecffe3846ff44feae0ff14d4e73bfa4,
title = "In Vitro Splicing Assays",
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.",
author = "Akila Mayeda and Krainer, {Adrian R.}",
year = "2012",
month = "2",
day = "2",
doi = "10.1002/9783527636778.ch30",
language = "English",
isbn = "9783527326068",
pages = "320--329",
booktitle = "Alternative pre-mRNA Splicing",
publisher = "Wiley-VCH",

}

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

In Vitro Splicing Assays. / Mayeda, Akila; Krainer, Adrian R.

Alternative pre-mRNA Splicing: Theory and Protocols. Wiley-VCH, 2012. p. 320-329.

Research output: Chapter in Book/Report/Conference proceedingChapter

TY - CHAP

T1 - In Vitro Splicing Assays

AU - Mayeda, Akila

AU - Krainer, Adrian R.

PY - 2012/2/2

Y1 - 2012/2/2

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84862885372&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862885372&partnerID=8YFLogxK

U2 - 10.1002/9783527636778.ch30

DO - 10.1002/9783527636778.ch30

M3 - Chapter

AN - SCOPUS:84862885372

SN - 9783527326068

SP - 320

EP - 329

BT - Alternative pre-mRNA Splicing

PB - Wiley-VCH

ER -

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