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A fluorimetric method for the estimation of G + C mol% content in microorganisms by thermal denaturation temperature
Abstract
Summary: 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.
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Cited by
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Human Infection with an Avian H9N2 Influenza A Virus in Hong Kong in 2003
Butt, K. M., Smith, G. J. D., Chen, H., Zhang, L. J., Leung, Y. H. C., Xu, K. M., Lim, W., Webster, R. G., Yuen, K. Y., Peiris, J. S. M., et al. (2005) · Journal of Clinical Microbiology · doi:10.1128/jcm.43.11.5760-5767.2005 · Cited by 594
Oberbeckmann, S., Loeder, M. G. J., Gerdts, G., & Osborn, A. M. (2014) · FEMS Microbiology Ecology · doi:10.1111/1574-6941.12409 · Cited by 540
Lopez-Siles, M., Khan, T. M., Duncan, S. H., Harmsen, H. J. M., Garcia-Gil, L. J., & Flint, H. J. (2012) · Applied and Environmental Microbiology · doi:10.1128/aem.06858-11 · Cited by 433
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Beatty, J. T., Overmann, J., Lince, M. T., Manske, A. K., Lang, A. S., Blankenship, R. E., Van Dover, C. L., Martinson, T. A., & Plumley, F. G. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0503674102 · Cited by 367
Metadata
- Authors
- Gonzalez, J. M., & Saiz‐Jimenez, C.
- Venue
- Environmental Microbiology
- Publisher
- Wiley
- Published
- 2002-11
- Volume
- 4
- Issue
- 11
- Pages
- 770-773
- ISSN
- 1462-2912 (ISSN-L)
- 1462-2920
- OpenAlex
- W2125054694
- Cited by
- 1,710
- References
- 0
- FWCI
- 13.07
- Type
- journal article
- Language
- en
- Open access
- closed
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- Gonzalez, J. M., & Saiz‐Jimenez, C. (2002). A fluorimetric method for the estimation of G + C mol% content in microorganisms by thermal denaturation temperature. Environmental Microbiology. https://doi.org/10.1046/j.1462-2920.2002.00362.x
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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