The DNeasy Plant system is designed for fast and easy purification of total cellular DNA (genomic, mitochondrial, chloroplast, viral) from plant cells and tissues or fungi. DNeasy purified DNA is suitable for all PCR, AFLP, RAPD, RFLP, and Southern-blotting applications.
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DNeasy Plant Kit Options |
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| Sample source | DNeasy Kit | Format |
| Plant tissues and cells or fungi: | ||
| Small-scale | DNeasy Plant Mini Kit | Mini spin columns |
| Large-scale | DNeasy Plant Maxi Kit | Maxi spin columns |
| High-throughput | DNeasy 96 Plant Kit | 96-well plates |
DNeasy Plant Kits use advanced silica-gel¨Cmembrane technology and simple spin procedures to isolate highly pure total cellular DNA from plant tissues and cells or fungi. DNeasy technology replaces cumbersome DNA isolation procedures such as CTAB, phenol, or chloroform extraction. Using the DNeasy procedure, alcohol precipitation is not necessary ¡ª purified DNA is ready for immediate use. Absorbance scans of DNeasy purified DNA show a symmetrical peak at 260 nm (see figure "DNA Purity Depends on Isolation Method"), confirming that the DNA is free of impurities, including enzyme inhibitors.
DNA Purity Depends on Isolation Method
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Spectrophotometric scans (220¨C320 nm) of DNA isolated from leaves/needles using the method of Dellaporta , CTAB , or the DNeasy Plant Mini Kit. Pure DNA typically shows a symmetrical peak at 260 nm and a smooth profile. Polysaccharides and other secondary metabolites, often copurified with plant DNA isolated using traditional methods, can interfere with OD readings (A260/A280), leading to errors in determination of concentration and purity. ¡¡ |
Samples are first mechanically disrupted and then chemically lysed. RNA is removed by RNase digestion during lysis. In the spincolumn procedure, cell debris is removed and samples are filtered and homogenized by centrifugation through a QIAshredder spin column. Buffering conditions are adjusted, precipitating proteins and polysaccharides, and the lysate is loaded onto the DNeasy Plant spin column or 96-well plate. During a brief spin, DNA selectively binds to the silica-gel membrane while contaminants pass through. Remaining contaminants and enzyme inhibitors are removed in one or two efficient wash steps. Pure DNA is then eluted in water or low-salt buffer, ready for use.
DNeasy purified DNA is sized up to 40 kb, with fragments of 20¨C25 kb predominating, and is suitable for applications such as: