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Updated: Jun 20, 2026

Quantification of dsDNA using the Hitachi F-7000 Fluorescence Spectrophotometer and PicoGreen Dye

Quantification of dsDNA using the Hitachi F-7000 Fluorescence Spectrophotometer and PicoGreen Dye

Published on: November 5, 2010

Metal-enhanced PicoGreen fluorescence: application for double-stranded DNA quantification.

A I Dragan¹, E S Bishop, J R Casas-Finet

¹Institute of Fluorescence, University of Maryland Biotechnology Institute, Baltimore, MD 21202, USA.

Analytical Biochemistry

|September 15, 2009

Summary

This summary is machine-generated.

PicoGreen (PG) dye enhances double-stranded DNA (dsDNA) detection sensitivity when combined with silver nanoparticles. This metal-enhanced fluorescence significantly amplifies dsDNA quantification capabilities.

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Area of Science:

Biochemistry
Nanotechnology
Analytical Chemistry

Background:

PicoGreen (PG) is a standard fluorescent probe for double-stranded DNA (dsDNA) detection.
Existing methods have limitations in sensitivity for dsDNA quantification.

Purpose of the Study:

To investigate the spectral properties of PicoGreen (PG) in proximity to silver nanoparticles.
To enhance the sensitivity of PG-based dsDNA detection and quantification.

Main Methods:

Studied spectral properties of free and DNA-bound PG with silver island films (SiFs).
Analyzed PG fluorescence at various concentrations (20 pM–3.5 µM) to confirm monomeric state.
Measured fluorescence enhancement due to metal-enhanced fluorescence (MEF) effect.

Main Results:

PG fluorescence increases ~1100-fold upon binding to dsDNA.
PG/dsDNA on SiFs shows a 7-fold increase due to MEF, totaling 7700-fold vs. free dye on glass.
Free PG on SiFs exhibited a 5-fold brightness decrease.
Total enhancement of ~38,000-fold achieved for PG on dsDNA on silver compared to free PG on SiFs.

Conclusions:

Silver nanoparticles significantly enhance PicoGreen fluorescence for dsDNA detection.
Metal-enhanced fluorescence offers a powerful method for amplified dsDNA quantification.