Ramalho Anabela S, Clarke Luka A, Amaral Margarida D
Faculty of Sciences, BioFiG-Centre for Biodiversity and Functional and Integrative Genomics, University of Lisboa, Lisboa, Portugal.
Methods Mol Biol. 2011;741:115-35. doi: 10.1007/978-1-61779-117-8_9.
Quantification and analysis of CFTR transcripts is of crucial importance not only for cystic fibrosis (CF) diagnosis and prognosis, but also in evaluating the efficiency of various therapeutic approaches to CF, including gene therapy. Reverse transcription (RT) followed by quantitative polymerase chain reaction (qPCR) is at present the most sensitive method for transcript abundance measurement. Classical RNA-based methods require significant expression levels in target samples for appropriate analysis, thus PCR-based methods have evolved towards reliable quantification. In this chapter we describe and discuss several protocols for the quantitative analysis of CFTR transcripts, including those variants that result from alternative splicing.
