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Quantification of dsDNA using the Hitachi F-7000 Fluorescence Spectrophotometer and PicoGreen Dye
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Rapid DNA quantification method in microplates using daunorubicine fluorescence quenching

M Gagne1, M Page

  • 1Department of Biological Chemistry, Faculty of Medicine, Universite Laval, Ste Foy, Quebec G1K 7P4, Canada.

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|May 9, 1998
PubMed
Summary

We developed a fast, affordable method to measure DNA using daunorubicin fluorescence quenching. This technique accurately quantifies DNA in crude extracts and PCR products, even with proteins present.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Accurate DNA quantification is crucial for molecular biology applications.
  • Existing methods can be time-consuming, expensive, or affected by contaminants like proteins.

Purpose of the Study:

  • To develop a rapid, sensitive, and inexpensive method for DNA quantification.
  • To assess the method's reliability in crude extracts and PCR products.
  • To evaluate potential interference from proteins.

Main Methods:

  • Utilized daunorubicin fluorescence quenching for DNA detection.
  • Developed a method applicable to crude DNA extracts and PCR reaction products.
  • Established a linear standard curve for DNA quantification.

Main Results:

  • Achieved a linear standard curve for DNA quantification ranging from 4.7 ng to 600 ng.
  • Demonstrated the method's sensitivity and speed.
  • Confirmed no significant interaction or interference from proteins.

Conclusions:

  • The daunorubicin fluorescence quenching method offers a rapid, sensitive, and cost-effective way to quantify DNA.
  • This technique is suitable for analyzing DNA in complex biological samples and PCR products.
  • The method's robustness in the presence of proteins enhances its practical utility.