For isolation of up to 30 µg total cellular DNA from plant cells and tissues, or fungi
Pure DNA, free from contaminants and enzyme inhibitors
Rapid isolation of ready-to-use DNA
No organic extraction, no ethanol precipitation
The DNeasy Plant Mini Kit provides fast and easy silica-based DNA purification in convenient spin column format. Typical yields are 3–30 μg of high-quality DNA, depending on the samples used. Purification of DNA using the DNeasy Plant Mini Kit can be automated on theQIAcube.
250 DNeasy Mini Spin Columns, 250 QIAshredder Mini Spin Columns, RNase A, Buffers, Collection Tubes (2 ml)
The DNeasy Plant Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Pure DNA (20–25 kb) for restriction analysis.
Agarose gel (0.8%) analysis of DNA isolated from the indicated leaves or needles with the DNeasy Plant Mini Kit was perfromed. Undigested DNA (0.5 µg, left) or 1 µg of EcoRI-digested DNA (right) were loaded in each pair of lanes. M: lambda-HindIII.
The DNeasy Plant Mini Kit allows rapid and efficient isolation of high-quality DNA from a wide variety of plant species and tissue types including the most demanding sources (see table "Selection of plant species processed with DNeasy Kits"). Samples may be fresh, frozen, or dried. The optimized DNeasy Plant procedure incorporates the QIAshredder spin column, a unique filtration and homogenization column that efficiently removes cell debris and improves sample handling following lysis.
Young leaves or needles (and other tissues, as indicated) were collected and immediately flash frozen. DNA isolation was then performed with the DNeasy Plant Mini Kit. 1Beechnut, 2dried leaves, 3callus, 4leaves from adult plant, 5endosperm, 6old leaves, rich in carbohydrates, 7buds. For more information on DNA isolation from other species including fungi, call QIAGEN Technical Services or your local distributor.
The typical yield is 3–30 µg, with a sample size of up to 100 mg wet weight, and an elution volume of 50–400 µl. The DNA yields vary between different species and tissues depending on genome size, ploidy, cell number, and age of tissue sample. Lower and higher range values correspond to arabidopsis and wheat, respectively.
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. Absorbance scans of DNeasy purified DNA show a symmetrical peak at 260 nm (see figures "DNA purity from oak leaves and pine needles"), confirming that the DNA is free of impurities, including enzyme inhibitors. DNeasy purified DNA is sized up to 40 kb (see figure "Pure DNA (20–25 kb) for restriction analysis"). Purified DNA can be used in a wide range of applications (see figures "PCR analysis" and "RAPD analysis").
DNeasy Plant Kits use advanced silica-membrane 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 cetyltrimethylammonium bromide (CTAB), phenol, or chloroform extraction. Using the DNeasy procedure, alcohol precipitation is not necessary — purified DNA is ready for immediate use. DNeasy sample preparation technology is fully licensed.
Samples are first mechanically disrupted and then chemically lysed (see flowchart "DNeasy Plant and DNeasy 96 Plant procedures"). RNA is removed by RNAse digestion during lysis. Cell debries, precipitated proteins, and polysaccharides are removed and the sample is homogenized by centrifugation through a QIAshredder spin column. Buffering conditions are adjusted and the lysate is loaded onto the DNEasy Plant Mini spin column. During a brief spin, DNA selectively binds to the silica 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.
The DNeasy Plant Mini Kit provides purification of DNA from plant tissue, including:
PCR, real-time PCR, blotting
Main sample type
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
2.5 ml (100 mg/ml; 7000 units/ml, solution)
Unit definition: That amount of enzyme causing the hydrolysis of RNA at a rate such that k (velocity constant) equals unity (Kunitz units) at 25°C and pH 5.0.