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.
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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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Harvey, S.E., Cheng, C. (2016). Methods for Characterization of Alternative RNA Splicing. In: Feng, Y., Zhang, L. (eds) Long Non-Coding RNAs. Methods in Molecular Biology, vol 1402. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3378-5_18
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DOI: https://doi.org/10.1007/978-1-4939-3378-5_18
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3378-5
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