从植物细胞、组织或真菌细胞样本中分离总DNA,产量可达260 µg

  • 高纯度DNA,无污染物和酶抑制剂
  • 快速纯化得到即用型DNA
  • 无需有机试剂抽提,无需乙醇沉淀

DNeasy Plant Maxi Kit通过基于硅胶膜的离心柱形式,可快速便捷的纯化DNA。常规产量为30–260 μg的高品质DNA,产量取决于样本来源。

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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)
68163
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The DNeasy Plant Maxi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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DNeasy Plant and DNeasy 96 Plant procedures.|PCR performance.|PCR analysis.|RAPD analysis.|DNA purity from oak leaves and pine needles.|
|DNA was isolated from arabidopsis leaves using either CTAB lysis (CTAB) or the DNeasy Plant Maxi Kit (DNeasy). Amplification reactions were prepared using purified DNA (1: 50 pg; 2: 100 pg) and primers to the Akin 10 gene. M: markers. (Data kindly provided by Alain Lecharny, Institut de Biotechnologie des Plantes, UMR CNRS-UPS Orsay, France.)|DNA (10 ng) from the indicated leaves or needles was amplified using universal primers for the noncoding intergenic spacer between the tRNA genes trnL (UAA) 5' exon and trnL (UAA) 3' exon of cpDNA (Taberlet, P. et al. (1991) Plant Mol. Biol. 17, 1105). M: 100 bp ladder.|DNA (50 ng) from leaves of the indicated in vitro-propagated sunflower (Helianthus) species was amplified using a 10-base RAPD primer and separated on a 1.5% agarose gel. M: 100 bp ladder. (Data kindly provided by H. J. Henn, Institute of Agricultural Botany, University of Bonn, Germany.)|Spectrophotometric scans (220–320 nm) of DNA isolated from pine 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.|
Performance

DNeasy Plant Maxi Kit可快速高效的纯化高品质DNA,可以从不同的植物种类和组织类型中进行纯化,包括较难处理的样本来源(参见下表)。样本可以是新鲜、冷冻或者干燥的。优化的DNeasy Plant操作流程与QIAshredder Maxi离心柱结合使用,独特的过滤和匀质柱,能够有效的去除细胞碎片并且优化裂解后的样本处理流程。

用DNeasy Kits处理的植物种类简表
Abies alba(银杉) Nicotiana tabacum(烟草)
Aesculus hippocastanum(七叶树) Oryza sativa(水稻)4
Arabidopsis thaliana(拟南芥) Pelargonium sp.(天竺葵)4
Avena sp.(燕麦) Petunia sp.4
Brassica napus(油菜) Pinus sylvestris(欧洲赤松),P. brutia5
Brassica oleracea(大头菜) Populus tremula(白杨)
Chicorium endivia(菊苣) Pseudotsuga menziesii(Douglas 冷杉)
Citrullini lanatus(西瓜) Quercus robur, Q. petrea(橡树)6,7
Egeria sp. Rhododendron sp.2,4
Fagus sylvatica(榉木)1 Rubus idaeus(覆盆子)
Helianthus spp.(向日葵) Solanum tuberosum(土豆)
Hordeum vulgare(大麦)2 Sphagnum palustre(苔)
Humulus sp.(酒花) Spinacia oleracea(菠菜)
Hydrilla sp. Taxus baccata(红豆杉)
Kalanchoe spp. Triticum aestivum(小麦)4
Lupinus sp. Ulmus glabra(榆树)6
Lycopersicon esculentum(番茄)3 Vitis spp.(葡萄)6
Myriophyllum sp. Zea mays(玉米)
幼叶或针叶(以及其他组织,如上)收集并立即速冻。使用DNeasy Plant Mini Kit进行DNA提取。1山毛榉、2干叶、3愈伤组织、4成年植物上的叶子、5胚乳、6富含碳水化合物的老树叶、7芽。包括真菌在内的其他物种的DNA提取的详细信息,请联系QIAGEN公司的技术支持或当地的经销商。

常规产量为30–260 µg,样本量至多1 g湿重,洗脱体积为500 µl至2 ml。DNA产量因种类和组织不同而变化,取决于基因组大小、倍体、细胞数量和组织样本的年龄。较低和较高的范围值分别对应于拟南芥和小麦。用DNeasy纯化的DNA进行吸光度扫描显示在260 nm处有一个对称峰(参见"DNA purity from pine needles"),证明DNA无酶抑制剂等杂质。纯化的DNA可用于多种应用(参见"PCR performance"、"PCR analysis和"RAPD analysis")。

Principle

DNeasy Plant Kits使用先进的硅胶膜技术和简单的离心操作流程从植物组织和细胞或者真菌中提取高纯度的细胞总DNA。DNeasy技术取代繁琐的DNA提取操作流程,如十六烷基三甲基溴化铵(CTAB)、苯酚或者氯仿萃取。使用DNeasy操作流程不需要乙醇沉淀,纯化的DNA可直接使用。DNeasy样本制备技术已经过认证。

Procedure

首先对样本进行机械搅拌,然后进行化学裂解(参见"DNeasy Plant and DNeasy 96 Plant procedures")。在裂解过程中,RNA被RNAase消化去除。细胞碎片、沉淀的蛋白质和多糖都被去除,并且样本离心通过QIAshredder Maxi离心柱实现匀浆。调节缓冲溶液条件,并将裂解产物上样到DNeasy Plant Maxi离心柱中。在短暂的离心过程中,DNA选择性结合到硅胶膜上,污染物通过。余下的污染物和酶抑制剂经过一或者两步高效的洗涤步骤去除。然后,用水或者低盐缓冲溶液将纯化的DNA洗脱,待用。

Applications

DNeasy Maxi Plant Kit可从植物组织中纯化DNA,包括:

  • 植物细胞
  • 植物组织
  • 真菌
Feature
Specifications
Applications PCR, real-time PCR, blotting
Elution volume 500 µl – 2 ml
Format Spin column
Main sample type Plant samples
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount <1 g
Technology Silica technology
Time per run or per prep <2 hours
Yield 30–260 µg

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Kit Handbooks
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The names of some of the buffers supplied with the kit have been changed. Please note that the composition of all the buffers remains unchanged.
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