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A Quick, Affordable Method for the Quantification of Gene Isoforms
Author(s) -
Bistulfi Gaia,
Sutherland Catherine
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.562.14
Subject(s) - gene isoform , gene , alternative splicing , biology , computational biology , genetics , primer (cosmetics) , gene expression , locus (genetics) , rna , rna splicing , chemistry , organic chemistry
Most genes are transcribed as several variants known as isoforms. Isoforms are transcripts from the same locus with either a different transcription start site or post‐transcriptional modifications, for example alternative splicing. As a result, isoforms might be characterized by different functions and/or stability. The differences between gene isoforms span from a few bases to several kilobases. In some cases, no region of the transcript might be specific to only one isoform, making quantification problematic. We devised a quick, reliable, one‐step method for the accurate quantification of gene isoforms, even in the latter case. We compared the isoforms' sequences using bioinformatic tools freely available online. We analyzed RNA samples with a one‐step real‐time RT‐PCR protocol, using primer sets specific to one or more isoforms and RNA standard curves as a reference. Differential analysis of the data provided reliable information on the copy number of each isoform in the original sample. This method might provide a first step in understanding the importance of differential expression of gene isoforms This work was supported by a D'Youville College research grant.