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Real Time RT-PCR02:57

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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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Related Experiment Video

Updated: May 1, 2026

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format
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Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format

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Clinical applications using digital PCR.

Francisco Bizouarn1

  • 1Gene Expression Division, Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA, 94547, USA, frank_bizouarn@Bio-Rad.com.

Methods in Molecular Biology (Clifton, N.J.)
|April 18, 2014
PubMed
Summary
This summary is machine-generated.

Molecular diagnostics enable personalized treatments by analyzing genetic profiles. Digital PCR (dPCR) offers accurate, rapid, and cost-effective detection of rare mutations and pathogens for improved patient care.

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Area of Science:

  • Molecular Diagnostics
  • Genomics
  • Biotechnology

Background:

  • Personalized medicine is shifting from "one-size-fits-all" to tailored treatments based on molecular profiling.
  • Genetic variations, such as in epidermal growth factor receptor (EGFR), can significantly impact treatment efficacy for cancers like lung and colorectal.
  • Ineffective treatments due to genetic mutations (e.g., BRAF V600E, KRAS G12/13) lead to increased costs and reduced patient well-being.

Purpose of the Study:

  • To highlight the growing importance of molecular diagnostics and patient-specific treatments.
  • To introduce digital PCR (dPCR) as a powerful technology for clinical diagnostics.
  • To discuss the optimization, validation, and applications of dPCR assays.

Main Methods:

  • Digital PCR (dPCR) assay optimization and validation.
  • Application of dPCR for pathogen/viral detection and quantitation.
  • Utilizing dPCR for copy number variation analysis and rare mutation detection.

Main Results:

  • dPCR demonstrates high specificity, accuracy, and speed, making it attractive for clinical diagnostics.
  • The technology is particularly impactful in pathogen detection, copy number variation analysis, and rare mutation abundance.
  • The QX100/QX200 platforms provide a foundation for dPCR assay development.

Conclusions:

  • Digital PCR is a transformative technology for molecular diagnostics.
  • dPCR facilitates more effective, personalized treatments by enabling precise detection of disease markers.
  • The expanding applications of dPCR promise to further enhance patient care and outcomes.