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Int. J. Mol. Sci. 2008, 9(11), 2306-2321; doi:10.3390/ijms9112306
Article

Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error

 and *
Received: 28 August 2008; in revised form: 19 November 2008 / Accepted: 24 November 2008 / Published: 25 November 2008
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Abstract: Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W.G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in plant tissue. We demonstrated that increasing the amount of plant material used for DNA extraction to 0.5-1.0 g considerably reduced sampling error and improved the reproducibility of DNA yield. The costs of DNA extraction at different scales and with different methods (commercial kits versus cetyltrimethylammonium bromide-based protocol) and qPCR systems (doubly labeled hybridization probes versus SYBR Green) were compared. A cost-effective protocol for the quantification of F. graminearum and F. culmorum DNA in wheat grain and maize stalk debris based on DNA extraction from 0.5-1.0 g material and real-time PCR with SYBR Green fluorescence detection was developed.
Keywords: Fusarium graminearum Schwabe; Fusarium culmorum W.G. Smith; realtime PCR; DNA extraction; sampling error; Fusarium head blight Fusarium graminearum Schwabe; Fusarium culmorum W.G. Smith; realtime PCR; DNA extraction; sampling error; Fusarium head blight
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Brandfass, C.; Karlovsky, P. Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error. Int. J. Mol. Sci. 2008, 9, 2306-2321.

AMA Style

Brandfass C, Karlovsky P. Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error. International Journal of Molecular Sciences. 2008; 9(11):2306-2321.

Chicago/Turabian Style

Brandfass, Christoph; Karlovsky, Petr. 2008. "Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error." Int. J. Mol. Sci. 9, no. 11: 2306-2321.


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