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Journal Article

Molecular quantification of genes encoding for green-fluorescent proteins

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Vandieken,  V.
Department of Biogeochemistry, Max Planck Institute for Marine Microbiology, Max Planck Society;

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Citation

Felske, A., Vandieken, V., Pauling, B. V., von Canstein, H. F., & Wagner-Dobler, I. (2003). Molecular quantification of genes encoding for green-fluorescent proteins. Journal of Microbiological Methods, 52(3), 297-304.


Cite as: http://hdl.handle.net/21.11116/0000-0001-D249-0
Abstract
A quantitative PCR approach is presented to analyze the amount of recombinant green fluorescent protein (gfp) genes in environmental DNA samples. The quantification assay is a combination of specific PCR amplification and temperature gradient gel electrophoresis (TGGE). Gene quantification is provided by a competitively coamplified DNA standard constructed by point mutation PCR. A single base difference was introduced to achieve a suitable migration difference in TGGE between the original target DNA and the modified standard without altering the PCR amplification efficiency. This competitive PCR strategy is a highly specific and sensitive way to monitor recombinant DNA in environments like the efflux of a biotechnological plant.