Methods for Characterization of Alternative RNA Splicing
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 wordsAlternative splicing RNA RT-PCR Variable exon Minigene Splicing factors Splicing regulation
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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