21. Quantification of Alternative Splice Variants
- Prof. Dr. Stefan Stamm1,
- Prof. Dr. Christopher W. J. Smith2,
- Prof. Dr. Reinhard Lührmann3
Published Online: 2 FEB 2012
DOI: 10.1002/9783527636778.ch21
Copyright © 2012 Wiley-VCH Verlag GmbH & Co. KGaA
Book Title
Alternative pre-mRNA Splicing: Theory and Protocols
Additional Information
How to Cite
Llorian, M. and Smith, C. W. J. (2012) Quantification of Alternative Splice Variants, in Alternative pre-mRNA Splicing: Theory and Protocols (eds S. Stamm, C. W. J. Smith and R. Lührmann), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527636778.ch21
Editor Information
- 1
University of Kentucky, Department of Molecular and Cellular Biochemistry, B278 Biomedical/Biological Sciences Research Building, 741 South Limestone Street, Lexington, KY 40536-0298, USA
- 2
University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QW, United Kingdom
- 3
Max Planck Institute for Biophysical Chemistry, Department of Cellular Biochemistry, Am Fassberg 11, 37077 Göttingen, Germany
Publication History
- Published Online: 2 FEB 2012
- Published Print: 11 JAN 2012
ISBN Information
Print ISBN: 9783527326068
Online ISBN: 9783527636778
- Summary
- Chapter
- References
Keywords:
- Quantitative RT-PCR;
- isoform ratio;
- alternative spliced variants;
- comparative quantitation
Summary
The increasing evidence that almost all human genes undergo alternative splicing highlights its importance as a central level of regulation of gene expression (see Chapters 3 Hertel, and 8 Smith). In order to characterize gene expression and to study splicing regulation, a robust, reliable and accurate method for the quantification of spliced isoforms is needed. Traditionally, several techniques have been used, including Northern blotting and nuclease protection, but overall RT-PCR has been the method of choice. In this chapter, the application of radiolabeled RT-PCR and real-time PCR to quantify splice variant changes will be described and discussed.