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 semiquantitative PCR methods. These methods require careful PCR primer design to ensure specific detection of particular splice isoforms. We will 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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Acknowledgments
This work was supported by research grants from the US National Institutes of Health (R01 CA182467, R35GM131876) to C.C. C.C. is a Cancer Prevention and Research Institute of Texas Scholar in Cancer Research (RR160009).
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Harvey, S.E., Lyu, J., Cheng, C. (2021). Methods for Characterization of Alternative RNA Splicing. In: Zhang, L., Hu, X. (eds) Long Non-Coding RNAs. Methods in Molecular Biology, vol 2372. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1697-0_19
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DOI: https://doi.org/10.1007/978-1-0716-1697-0_19
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