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Related Experiment Videos

Competitive PCR for precise nucleic acid quantification.

Lorena Zentilin1, Mauro Giacca

  • 1Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano 99, Trieste, Italy.

Nature Protocols
|September 15, 2007
PubMed
Summary
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Accurately quantify nucleic acids using competitive PCR, a method that coamplifies target and competitor DNA. This technique offers superior accuracy for absolute DNA quantification compared to other PCR methods.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Precise quantification of nucleic acids is crucial for experimental and diagnostic applications.
  • Existing quantitative PCR methods have limitations in accuracy.

Purpose of the Study:

  • To present a comprehensive protocol for competitive PCR and RT-PCR.
  • To enable accurate absolute quantification of target DNA molecules.

Main Methods:

  • Design and construction of competitor DNA molecules.
  • Coamplification of target and competitor DNA using specific primers.
  • Post-PCR analysis for data interpretation and quantification.

Main Results:

  • Competitive PCR allows for precise quantification of absolute target DNA amounts.

Related Experiment Videos

  • This method demonstrates higher accuracy than real-time PCR.
  • The protocol can be completed within 24 hours once primers are available.

    • Conclusions:

    • Competitive PCR provides a highly accurate method for nucleic acid quantification.
    • This protocol facilitates reliable DNA quantification in research and diagnostics.
    • The method is robust against variations affecting PCR amplification.