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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae
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Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae

Published on: March 10, 2012

Quantification of CFTR transcripts.

Anabela S Ramalho1, Luka A Clarke, Margarida D Amaral

  • 1Faculty of Sciences, BioFiG-Centre for Biodiversity and Functional and Integrative Genomics, University of Lisboa, Lisboa, Portugal. asramalho@fc.ul.pt

Methods in Molecular Biology (Clifton, N.J.)
|May 20, 2011
PubMed
Summary
This summary is machine-generated.

Accurately quantifying cystic fibrosis transmembrane conductance regulator (CFTR) transcripts is vital for diagnosing cystic fibrosis and assessing gene therapy effectiveness. This chapter details sensitive reverse transcription quantitative polymerase chain reaction (RT-qPCR) protocols for CFTR transcript analysis.

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Last Updated: Jun 1, 2026

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14:56

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15:12

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Accurate quantification of cystic fibrosis transmembrane conductance regulator (CFTR) transcripts is essential for cystic fibrosis (CF) diagnosis, prognosis, and evaluating therapeutic strategies like gene therapy.
  • Traditional RNA analysis methods necessitate substantial expression levels, driving the evolution of sensitive PCR-based techniques for reliable transcript abundance measurement.

Purpose of the Study:

  • To describe and discuss protocols for the quantitative analysis of CFTR transcripts.
  • To include methods for analyzing CFTR variants arising from alternative splicing.

Main Methods:

  • Utilizing reverse transcription (RT) followed by quantitative polymerase chain reaction (qPCR) as the primary sensitive method for transcript abundance measurement.
  • Detailing specific protocols for the quantitative analysis of CFTR transcripts.

Main Results:

  • The described RT-qPCR protocols enable sensitive and reliable quantification of CFTR transcript levels.
  • The methods discussed are applicable to analyzing CFTR variants, including those produced by alternative splicing.

Conclusions:

  • Quantitative analysis of CFTR transcripts using RT-qPCR is crucial for advancing cystic fibrosis research and clinical applications.
  • The presented protocols offer a sensitive and reliable approach for measuring CFTR transcript abundance and its variants.