从植物组织分离细胞内总DNA,每孔至多15 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

DNeasy 96 Plant Kit采用基于硅胶膜技术的离心柱和96孔板方式快速方便的纯化DNA。一般产量为1–15 µg高品质DNA,取决于植物样本的类型。

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DNeasy 96 Plant Kit (6)
For 6 x 96 DNA minipreps: 6 DNeasy 96 Plates, Buffers, Reagents, RNase A, S-Blocks, Collection Microtubes (1.2 ml), Caps, AirPore Tape Sheets
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The DNeasy 96 Plant 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 procedure.|Reproducible DNA yield and quality.|Reliable PCR.|DNA purity from pine needles.|DNeasy 96 Plant plate.|Uniformity of DNA yields from 96 wheat samples.|
|Genomic DNA was purified from different plant species following disruption in liquid nitrogen. DNA was eluted in 2 x 100 µl Buffer AE. Eluates (10 µl) were subjected to gel electrophoresis. Average yields are given under the plant names. M: lambda–HindIII.|Genomic DNA purified from different plant species was amplified by PCR. Eluate (5 µl) was used as template, and 30 µl of each PCR product were loaded per lane. M: markers.|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.||Ninety-six leaf samples were collected from a batch of wheat plants grown under identical conditions. Samples (50 mg) were disrupted in liquid nitrogen and DNA was isolated using the DNeasy 96 Plant procedure.
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Performance

DNeasy 96 Plant Kit可在2个小时内从96或192个植物叶片组织样本中高通量纯化高品质DNA。简单的96孔操作流程能高重复的得到高产量的细胞内总DNA(参见"Reproducible DNA yield and quality")。一般产量为每50 mg植物叶片组织得到1–15 µg DNA,用100–200 µl洗脱液洗脱。DNA产量因种类不同而不同,取决于基因组大小、倍体、细胞数量和组织样本的年龄。

用DNeasy纯化得到的DNA至多长40 kb,以20–25 kb的片段为主。可有效地去除多糖、蛋白质和酶抑制剂。DNeasy纯化得到的DNA吸光度扫描显示在260 nm处有对称的峰,说明DNA无杂质,包括酶抑制剂(参见"DNA purity from pine needles")。纯化得到的DNA用于多种下游应用(参见"Reliable PCR")。

裂解高通量样本

QIAGEN的商业化高通量裂解体系TissueLyser,仅在2–4分钟内就可快速高效的裂解192个植物组织样本。TissueLyser统一裂解样本,可获得可重复的高品质结果,用于下游应用如从植物组织中分离基因组DNA。

Principle

DNeasy 96 Plant Kit采用先进的硅胶膜技术和简单的离心柱操作流程,从植物组织和细胞或真菌中分离高纯度细胞内总DNA。精简的DNeasy 96操作流程和方便的96孔方式无需繁琐的标记、一次一步的移液操作和DNA重悬浮步骤,有助于较少交叉感染。DNeasy技术代替了繁琐的DNA分离操作流程如十六烷基三甲基溴化铵(CTAB)、苯酚或氯仿提取。采用DNeasy操作流程,无需乙醇沉淀,且纯化得到的DNA可直接应用。DNeasy样本制备技术已获得全部许可。

Procedure

样本首先经机械裂解,然后经化学裂解(参见"DNeasy Plant procedure")。裂解过程中通过RNA酶消化去除RNA。细胞沉淀、蛋白沉淀和多糖可通过离心去除。调节缓冲条件,随后将裂解物上样到DNAeasy Plant 96孔板上(参见"DNeasy 96 Plant plate")。在短时离心过程中,DNA可选择性结合到硅胶膜上,而污染物通过。余下的污染物和酶抑制剂可通过一步有效的洗涤步骤除去。然后用纯水或低盐缓冲液洗脱纯DNA待用。处理DNeasy 96孔板需要使用QIAGEN 96-Well-Plate Centrifugation System。

Applications

DNAeasy 96 Plant Kit能从植物组织中纯化得到DNA,包括:

  • 植物细胞
  • 植物组织
  • 真菌
Feature
Specifications
Applications PCR, real-time PCR, blotting
Elution volume 100–200 µl
Format 96-well plate
Main sample type Plant samples
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount 50 mg
Technology Silica technology
Time per run or per prep <2 hours (192 samples)
Yield 1–15 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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