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DNeasy Plant Mini Kit
植物细胞，组织或真菌细胞样品分离总DNA,最高产量可达 30 µg
纯DNA，无污染物和酶抑制剂
快速纯化得到即用型DNA
无需有机抽提，无需乙醇沉淀
Format: Mini spin columns
with 2 ml collection tubes
Sample source: Plant cells and tissues, fungi
Sample size: Up to 100 mg wet weight
Preparation time: <1 hour
Typical yield:* 3–30 µg
Elution volume: 50–400 µl
Procedure
Samples are first mechanically disrupted and then chemically lysed. RNA is removed by RNase digestion during lysis. 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. 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 (see flowchart "DNeasy Plant and DNeasy 96 Plant Procedures").
Downstream applications
The DNeasy Plant procedure yields pure nucleic acid, free of polysaccharides and other secondary metabolites often copurified using conventional methods. Such impurities can interfere with spectrophotometric readings and inhibit enzymatic reactions. DNeasy purified DNA is sized up to 40 kb (see figure "Pure DNA (20–25 kb) for Restriction Analysis"), and is suitable for downstream applications such as:
PCR (see figure "PCR Analysis of DNA from Different Plant Species")
AFLP
RFLP
RAPD (see figure "RAPD Analysis of Sunflower Species")
Southern blotting
Microsatellite analysis
Real-time PCR
Cited References
Dellaporta, S.L. (1983) A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1, 19.
Doyle, J.J. and Doyle J.L. (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19, 11. |