title: Methods for Characterization of Alternative RNA Splicing.
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.
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Samuel E
authors.initials: SE
authors.lastname: Harvey
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA. chengc@northwestern.edu.
authors.forename: Chonghui
authors.initials: C
authors.lastname: Cheng
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D005091:Exons; D057809:HEK293 Cells; D006801:Humans; D012313:RNA / Q000235:genetics*; D015347:RNA Probes / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
chemical_list: D015347:RNA Probes; D012313:RNA
keywords: Alternative splicing; Minigene; RNA; RT-PCR; Splicing factors; Splicing regulation; Variable exon