A fluorimetric method for the estimation of G+C mol% content in microorganisms by thermal denaturation temperature
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- 1 November 2002
- journal article
- research article
- Published by Wiley in Environmental Microbiology
- Vol. 4 (11), 770-773
- https://doi.org/10.1046/j.1462-2920.2002.00362.x
Abstract
G+C mol% content in microorganisms is one of the recommended characteristics for the standard description of bacterial species. In this study we present a novel fluorimetric method to estimate the G+C mol% content in microorganisms. Double‐stranded DNA was specifically stained with SYBR Green I, and its thermal denaturalization was followed by measuring a decrease in fluorescence using a real‐time PCR thermocycler. Unlike most previous determinations of G+C mol%, in this study only DNA from microorganisms with an available completely sequenced genome were used to prepare the calibration curves. Calibration curves showed a linear relationship between G+C mol% content and melting temperature and they were performed both in the absence and presence of 30% formamide. This protocol proves to be a rapid and inexpensive method to estimate DNA base ratios of novel microorganisms.Keywords
This publication has 10 references indexed in Scilit:
- Pyrococcus horikoshii sp. nov., a hyperthermophilic archaeon isolated from a hydrothermal vent at the Okinawa TroughExtremophiles, 1998
- Polyphasic taxonomy, a consensus approach to bacterial systematics.Microbiological Reviews, 1996
- Extraction, Purification, and Analysis of DNA from Soil BacteriaPublished by Springer Science and Business Media LLC ,1995
- Precise Measurement of the G+C Content of Deoxyribonucleic Acid by High-Performance Liquid ChromatographyInternational Journal of Systematic and Evolutionary Microbiology, 1989
- Report of the Ad Hoc Committee on Reconciliation of Approaches to Bacterial SystematicsInternational Journal of Systematic and Evolutionary Microbiology, 1987
- Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100°CArchiv für Mikrobiologie, 1986
- Determination of DNA base composition by reversed-phase high-performance liquid chromatographyFEMS Microbiology Letters, 1984
- The Quantitative Measurement of DNA Hybridization from Renaturation RatesEuropean Journal of Biochemistry, 1970
- Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperatureJournal of Molecular Biology, 1962
- A procedure for the isolation of deoxyribonucleic acid from micro-organismsJournal of Molecular Biology, 1961