For isolation of up to 260 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 Maxi Kit provides fast and easy silica-based DNA purification in spin column format. Typical yields are 30–260 μg of high-quality DNA, depending on the sample source.
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DNeasy Plant Maxi Kit (24)
24 DNeasy Maxi Spin Columns, 24 QIAshredder Maxi Spin Columns, RNase A, Buffers, Collection Tubes (50 ml)
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68163
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DNeasy Plant Maxi Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。
DNeasy Plant和DNeasy 96 Plant实验流程。|PCR表现。|PCR分析。|RAPD分析。|从松针中纯化获得的DNA纯度。|
|使用CTAB裂解液(CTAB)或DNeasy Plant Maxi Kit (DNeasy)从拟南芥叶片中纯化DNA。使用纯化的NA(1: 50 pg; 2: 100 pg)和Akin 10基因的引物进行扩增反应。. M:标记物。(数据由法国Alain Lecharny, Institut de Biotechnologie des Plantes, UMR CNRS-UPS Orsay提供)|使用tRNA基因trnL(UAA)的5'外显子和cpDNA的trnL(UAA)3'外显子间的非编码基因间隔的通用引物,扩增图示叶片或松针的DNA(10 ng)。(Taberlet, P. et al. (1991) Plant Mol. Biol. 17, 1105)。M: 100 bp ladder。|使用10-base RAPD引物对体外培植的向日葵(Helianthus)叶片的DNA(50 ng)进行扩增,然后在1.5%的琼脂糖凝胶上进行分离。M: 100 bp ladder。(数据由德国University of Bonn,Institute of Agricultural Botany的H. J. Henn提供)。|使用Dellaporta、CTAB、或DNeasy Plant Mini Kit从松针中纯化DNA,对DNA进行分光光度法扫描(220–320 nm)。纯DNA通常在260 nm处出现对称峰,且较为平滑。使用传统纯化方法时,多糖和其他次生代谢产物经常与植物DNA一同被分离,会干扰OD读数(A260/A280),导致对DNA浓度和纯度的错误分析。|
性能
The DNeasy Plant Maxi 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 Maxi spin column, a unique filtration and homogenization column that efficiently removes cell debris and improves sample handling following lysis.
| Abies alba (silver fir) |
Nicotiana tabacum (tobacco) |
| Aesculus hippocastanum (horse chestnut) |
Oryza sativa (rice)4 |
| Arabidopsis thaliana (thale cress) |
Pelargonium sp. (geranium)4 |
| Avena sp. (oat) |
Petunia sp.4 |
| Brassica napus (oilseed rape) |
Pinus sylvestris (Scotch pine), P. brutia5 |
| Brassica oleracea (kohlrabi) |
Populus tremula (aspen) |
| Chicorium endivia (chicory) |
Pseudotsuga menziesii (Douglas fir) |
| Citrullini lanatus (water melon) |
Quercus robur, Q. petrea (oak)6,7 |
| Egeria sp. |
Rhododendron sp.2,4 |
| Fagus sylvatica (beech)1 |
Rubus idaeus (raspberry) |
| Helianthus spp. (sunflower) |
Solanum tuberosum (potato) |
| Hordeum vulgare (barley)2 |
Sphagnum palustre (moss) |
| Humulus sp. (hops) |
Spinacia oleracea (spinach) |
| Hydrilla sp. |
Taxus baccata (yew) |
| Kalanchoe spp. |
Triticum aestivum (wheat)4 |
| Lupinus sp. |
Ulmus glabra (elm)6 |
| Lycopersicon esculentum (tomato)3 |
Vitis spp. (grape)6 |
| Myriophyllum sp. |
Zea mays (maize) |
The typical yield is 30–260 µg, with a sample size of up to 1 g wet weight, and an elution volume of 500 µl to 2 ml. 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. 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. Purified DNA can be used in a wide range of applications (see figures "PCR performance", "PCR analysis", and "RAPD analysis").
原理
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.
操作流程
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 Maxi spin column. Buffering conditions are adjusted and the lysate is loaded onto the DNeasy Plant Maxi 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.
应用
The DNeasy Plant Maxi Kit provides purification of DNA from plant tissue, including:
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Plant cells
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Plant tissues
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Fungi
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特点
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参数
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应用
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PCR, real-time PCR, blotting
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洗脱体积
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500 µl – 2 ml
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规格
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Spin column
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主要样本类型
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Plant samples
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处理
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Manual
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纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白
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DNA
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样本量
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<1 g
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技术
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Silica technology
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每次运行或制备时间
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<2 hours
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产量
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30–260 µg
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FAQ ID -12
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浏览
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FAQ ID -114
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浏览
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FAQ ID -616
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浏览
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FAQ ID -728
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浏览
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FAQ ID -730
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浏览
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FAQ ID -754
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浏览
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FAQ ID -305
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浏览
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图片
DNeasy Plant和DNeasy 96 Plant实验流程。
PCR表现。
使用CTAB裂解液(CTAB)或DNeasy Plant Maxi Kit (DNeasy)从拟南芥叶片中纯化DNA。使用纯化的NA(1: 50 pg; 2: 100 pg)和Akin 10基因的引物进行扩增反应。. M:标记物。(数据由法国Alain Lecharny, Institut de Biotechnologie des Plantes, UMR CNRS-UPS Orsay提供)
PCR分析。
使用tRNA基因trnL(UAA)的5'外显子和cpDNA的trnL(UAA)3'外显子间的非编码基因间隔的通用引物,扩增图示叶片或松针的DNA(10 ng)。(Taberlet, P. et al. (1991) Plant Mol. Biol. 17, 1105)。M: 100 bp ladder。
RAPD分析。
使用10-base RAPD引物对体外培植的向日葵(Helianthus)叶片的DNA(50 ng)进行扩增,然后在1.5%的琼脂糖凝胶上进行分离。M: 100 bp ladder。(数据由德国University of Bonn,Institute of Agricultural Botany的H. J. Henn提供)。
从松针中纯化获得的DNA纯度。
使用Dellaporta、CTAB、或DNeasy Plant Mini Kit从松针中纯化DNA,对DNA进行分光光度法扫描(220–320 nm)。纯DNA通常在260 nm处出现对称峰,且较为平滑。使用传统纯化方法时,多糖和其他次生代谢产物经常与植物DNA一同被分离,会干扰OD读数(A260/A280),导致对DNA浓度和纯度的错误分析。
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