LEM-PCR: a method for determining relative transcript isoform proportions using real-time PCR without a standard curve.

ABSTRACT: Many genes express multiple transcript isoforms generated by alternative splicing of mRNA. Using real-time PCR, it is straightforward to determine the relative expression level of each isoform independently. However, it is less trivial to determine the relative proportions of different isoforms in a cDNA sample. The relative proportions of different isoforms can be important, as a small change in a highly abundant transcript may be more relevant than a large change in a minimally expressed transcript. Currently, determining the relative proportions of isoforms requires the construction of a standard curve using recombinant plasmid DNA or genomic DNA. As recombinant or genomic DNA standards often amplify with different efficiencies to cDNA samples, they may give under- or overestimations of

SUBMITTER: Virtue S 

PROVIDER: S-EPMC4289856 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature