Abstract
Real-time polymerase chain reaction (PCR) is a molecular biology technique based on the detection of the fluorescence produced by a reporter molecule, which increases as the reaction proceeds proportionally to the accumulation of the PCR product within each amplification cycle. The fluorescent reporter molecules include dyes that bind to the double-stranded DNA (i.e., SYBR® Green) or sequence-specific probes (i.e., Molecular Beacons or TaqMan® Probes). Real-time PCR provides a tool for accurate and sensitive quantification of target fungal DNA. Here, we describe a TaqMan real-time PCR method for specific detection and quantification of Alternaria spp. The method uses Alternaria-specific primers and probe, targeting the internal transcribed spacer regions ITS1 and ITS2 of the rRNA gene, and a positive amplification control based on 18S rRNA gene.
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Acknowledgments
This work was supported by Grant No. AGL 2006-07659 from the Ministerio de Educación y Ciencia of Spain and the Programa de Vigilancia Sanitaria 2009/AGR-1489 from the Comunidad de Madrid (Spain).
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Pavón, M.Á., López-Calleja, I.M., González, I., Martín, R., García, T. (2017). Targeting Conserved Genes in Alternaria Species. In: Moretti, A., Susca, A. (eds) Mycotoxigenic Fungi. Methods in Molecular Biology, vol 1542. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6707-0_6
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DOI: https://doi.org/10.1007/978-1-4939-6707-0_6
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