Quantitative methods for the analysis of CFTR transcripts/splicing variants

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Abstract

In cystic fibrosis (CF), transcript analysis and quantification are important for diagnosis, prognosis and also as surrogate markers for some therapies including gene therapy. Classical RNA-based methods require significant expression levels in target samples for appropriate analysis, thus PCR-based methods are evolving towards reliable quantification. Various protocols for the quantitative analysis of CFTR transcripts (including those resulting from splicing variants) are described and discussed here.

Keywords

Transcripts
RT-PCR
Quantitative PCR
Real-time PCR
Splicing
Alternative splicing

Abbreviations

CERES
Composite Exonic Regulatory Element of Splicing
ddNTPs
dideoxynucleotides
NBD
nucleotide-binding domain
OAT
ornithine amino transferase
PCR
polymerase chain reaction
RT
reverse transcriptase
SAB
sum above background

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