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Published on: November 15, 2017
A simple DNA extraction method suitable for PCR detection of genetically modified maize.
Manuel Porcar1, Silvia Ramos, Amparo Latorre
1Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de València, Apartado Postal 22085, 46071 València, Spain.
Journal of the Science of Food and Agriculture
|September 14, 2010
Summary
A rapid, NaOH-based DNA extraction method allows for quick isolation of DNA from maize tissues. This method enables sensitive polymerase chain reaction (PCR) detection of transgenic DNA, including event MON810.
Area of Science:
- Agricultural Biotechnology
- Molecular Biology
Background:
- Polymerase chain reaction (PCR) is vital for detecting transgenic DNA.
- Accurate PCR requires pure DNA, free from inhibitors like plant polysaccharides.
- Existing DNA extraction methods can be time-consuming and destructive.
Purpose of the Study:
- To develop a fast and simple DNA extraction method for maize.
- To isolate DNA suitable for PCR-based detection of transgenic event MON810.
Main Methods:
- A sodium hydroxide (NaOH)-based DNA extraction protocol was developed.
- The method utilizes small amounts of maize leaf tissue or single kernels.
- DNA-containing solutions were used as templates for PCR.
Main Results:
- The NaOH-based method is time-saving, taking only 5 minutes.
- DNA extraction is possible from small leaf portions (1 mg) or single kernels.
- Reproducible detection of transgenic DNA was achieved using PCR.
Conclusions:
- The procedure offers a quick extraction step and non-destructive kernel testing.
- Robust PCR detection of transgenic DNA (event MON810) is enabled.
- Highly specific amplicons are generated using MON810-specific oligonucleotides.
