A reliable method for quantification of splice variants using RT-qPCR.

ABSTRACT:

Background

The majority of protein isoforms arise from alternative splicing of the encoding primary RNA transcripts. To understand the significance of single splicing events, reliable techniques are needed to determine their incidence. However, existing methods are labour-intensive, error-prone or of limited use.

Results

Here, we present an improved method to determine the relative incidence of transcripts that arise from alternative splicing at a single site. Splice variants were quantified within a single sample using one-step reverse transcription quantitative PCR. Amplification products obtained with variant specific primer pairs were compared to those obtained with primer pairs common to both variants. The identities of variant specific amplicons were simultaneously ver

SUBMITTER: Camacho Londono J 

PROVIDER: S-EPMC4793508 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature