Related Experiment Video
Updated: Jul 19, 2026

Studying DNA Looping by Single-Molecule FRET
Published on: June 28, 2014
A simple fluorimetric method for the estimation of DNA-DNA relatedness between closely related microorganisms by
Juan M Gonzalez1, Cesareo Saiz-Jimenez
1Instituto de Recursos Naturales y Agrobiología, CSIC, P.O. Box 1052, 41080 Sevilla, Spain. jmgrau@irnase.csic.es
Abstract:
Determination of whole-genome DNA-DNA similarity is today a standard technique for species delineation in microbial taxonomy. However, these studies demand hard-to-perform and time-consuming experiments. Herein, we present an easy and rapid fluorimetric method to estimate DNA-DNA relatedness between microbial strains from differences of the thermal denaturation temperatures of hybrid and homologous genomic DNA. Double-stranded DNA was specifically stained with SYBR Green I, and its thermal denaturalization was followed by measuring a decrease in fluorescence. A quantitative, real-time PCR thermocycler was used to perform the experiment and obtain fluorescence determinations at increasing temperatures. The proposed method was validated by comparing species of the hyperthermophilic genera Pyrococcus and Thermococcus. The method proves to be an easy, rapid, and inexpensive alternative to estimate DNA-DNA relatedness between closely related species.
Related Concept Videos
DNA Isolation
Real Time RT-PCR
The real-time quantification of the number of amplified products is...
Modern Molecular Taxonomy
Methods to Assess Microbial Populations

