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Methods for Characterization of Alternative RNA Splicing

  • Samuel E. Harvey
  • Chonghui ChengEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1402)

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.

Key words

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

Notes

Acknowledgement

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.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Division of Hematology/Oncology, Department of Medicine, Robert H. Lurie Comprehensive Cancer CenterNorthwestern University Feinberg School of MedicineChicagoUSA

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