Methods for characterization of alternative RNA splicing

Samuel E. Harvey; Chonghui Cheng

doi:10.1007/978-1-4939-3378-5_18

Methods for characterization of alternative RNA splicing

Samuel E. Harvey, Chonghui Cheng

Medicine, Hematology Oncology Division

Research output: Chapter in Book/Report/Conference proceeding › Chapter

40 Scopus citations

Abstract

Quantification of alternative splicing to detect the abundance of differentially spliced isoforms of a gene in total RNA can be accomplished via RT-PCR using both quantitative real-time and semi-quantitative PCR methods. These methods require careful PCR primer design to ensure specific detection of particular splice isoforms. We also describe analysis of alternative splicing using a splicing “minigene” in mammalian cell tissue culture to facilitate investigation of the regulation of alternative splicing of a particular exon of interest.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	229-241
Number of pages	13
DOIs	https://doi.org/10.1007/978-1-4939-3378-5_18
State	Published - Jan 1 2016

Publication series

Name	Methods in Molecular Biology
Volume	1402
ISSN (Print)	1064-3745

Funding

This work was supported by research grants from the US National Institutes of Health (R01GM110146) and the American Cancer Society (RSG-09-252-01-RMC) to C.C.

Keywords

Alternative splicing
Minigene
RNA
RT-PCR
Splicing
Splicing factors
Variable exon
regulation

ASJC Scopus subject areas

Genetics
Molecular Biology

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10.1007/978-1-4939-3378-5_18

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KW - Minigene

KW - RNA

KW - RT-PCR

KW - Splicing

KW - Splicing factors

KW - Variable exon

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Methods for characterization of alternative RNA splicing

Abstract

Publication series

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Keywords

ASJC Scopus subject areas

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