A simple fluorimetric method for the estimation of DNA-DNA relatedness between closely related microorganisms by thermal denaturation temperatures

Extremophiles : Life Under Extreme Conditions
Juan M Gonzalez, Cesareo Saiz-Jimenez

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.

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Citations

Aug 31, 2013·Antonie van Leeuwenhoek·Patrycja GolinskaMichael Goodfellow
Feb 2, 2013·Antonie van Leeuwenhoek·Patrycja GolinskaMichael Goodfellow
May 30, 2013·Antonie van Leeuwenhoek·Patrycja GolinskaMichael Goodfellow
Aug 7, 2010·International Journal of Systematic and Evolutionary Microbiology·Byung-Yong KimMichael Goodfellow
Mar 2, 2011·Canadian Journal of Microbiology·Jennifer Loveland-CurtzeJean E Brenchley
Mar 19, 2013·Antonie van Leeuwenhoek·Leonardo José da SilvaTiago Domingues Zucchi
Jul 24, 2014·Archives of Microbiology·Syed G DastagerWen-Jun Li
Nov 27, 2014·Antonie van Leeuwenhoek·Demet Tatar, Nevzat Sahin
Nov 30, 2014·Antonie van Leeuwenhoek·Vidya V SonalkarSyed G Dastager
Sep 4, 2010·Current Microbiology·William J Hunter, Daniel K Manter
Nov 26, 2010·Current Microbiology·Daniel K ManterJorge M Vivanco
Oct 18, 2015·International Journal of Systematic and Evolutionary Microbiology·B DivyasreeCh V Ramana
Dec 17, 2015·Antonie van Leeuwenhoek·Aysel VeyisogluNevzat Sahin
Nov 9, 2015·Antonie van Leeuwenhoek·Fadime Ozdemir-KocakNevzat Sahin
Oct 3, 2015·Antonie van Leeuwenhoek·Nevzat SahinMichael Goodfellow
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