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Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR
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Quantitative DNA Analysis Using Droplet Digital PCR.

Rolf H A M Vossen1, Stefan J White2

  • 1Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2016
PubMed
Summary
This summary is machine-generated.

Droplet digital PCR (ddPCR) precisely quantifies DNA by partitioning samples into thousands of droplets for amplification. This method enables accurate genotyping of copy number variations and quantification of sequencing libraries.

Keywords:
Copy number variationDNA quantitationDigital PCRNGS

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Droplet digital PCR (ddPCR) offers precise nucleic acid quantification.
  • It relies on partitioning individual DNA molecules into separate droplets for amplification.

Purpose of the Study:

  • To outline the fundamental principles of ddPCR.
  • To demonstrate ddPCR applications in genotyping copy number variation (CNV) and quantifying Illumina sequencing libraries.

Main Methods:

  • Utilizing the Bio-Rad QX200 system for ddPCR.
  • Compartmentalizing individual DNA molecules within droplets for amplification.
  • Detecting amplified product via fluorescent signals to determine positive droplets.

Main Results:

  • Successful application of ddPCR for accurate copy number variation genotyping.
  • Effective quantification of Illumina sequencing libraries using ddPCR.
  • Demonstrated precision and reliability of the ddPCR technique.

Conclusions:

  • ddPCR is a powerful tool for precise DNA quantification.
  • The described applications highlight ddPCR's versatility in genetic analysis and library preparation.