从凝胶和溶液中纯化DNA片段(40 bp–50 kb)

  • 高效回收40 bp到50 kb的DNA
  • 从TAE或TBE琼脂糖凝胶和聚丙烯酰胺凝胶中回收DNA
  • 无碘化钠,避免干扰后续反应
  • 避免DNA大片段的剪切

QIAEX II Gel Extraction Kit提供硅胶颗粒悬浮液,在离液盐存在的条件下,DNA片段结合到硅胶颗粒上。QIAEX II Suspension添加到溶液或溶解的琼脂糖凝胶中结合DNA。通过简单的离心操作收集硅胶颗粒,洗涤后用Tris缓冲液或纯水洗脱得到40 bp–50 kb的DNA。

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QIAEX II Gel Extraction Kit (150)
For 150 extractions: 3 x 0.5 ml QIAEX II Suspension, Buffers
20021
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QIAEX II Gel Extraction Kit (500)
For 500 extractions: 5 x 1.0 ml QIAEX II Suspension, Buffers
20051
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The QIAEX II Gel Extraction 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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QIAEX II procedure.|pH Indicator Dye.|Consistent recovery.|
|pH indicator dye in the solubilization and binding buffer allows easy visual determination of optimal pH for DNA adsorption (pH ≤7.5). An incorrect binding-mixture pH can occur if the agarose gel electrophoresis buffer was frequently used or incorrectly prepared. In this case, the pH can be easily adjusted by addition of 10 µl 3 M sodium acetate, pH 5.0.
|Recovery of various amounts of a 2.9 kb DNA fragment from 1% agarose gels using the QIAEX II Gel Extraction Kit is shown.
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Performance

使用QIAEX II Gel Extraction Kit可高效回收获得10 ng到10 µg DNA(参见"Consistent recovery")。使用30 µl的QIAEX II悬浮液,多功能的批处理程序使结合能力很容易达到15 µg的DNA。

QIAEX II Gel Extraction Kit应用硅颗粒纯化DNA片段(40 bp–50 kb),回收率可达60–95%。10 µl QIAEX II悬浮液可结合至多5 µg的DNA,后续洗脱液只需20 µl。

根据片段大小回收
DNA大小回收率,百分比*
44 bp 75
75 bp 75
500 bp 95
7.5 kb 85
23.5 kb 75
48.5 kb 60
* 从上样2 µg的1% TAE琼脂糖凝胶回收
Principle

使用QIAEX II体系纯化DNA片段的基本条件是,在离液盐存在的情况下,琼脂糖处于溶解状态,并且核酸可选择性的吸附到QIAEX II硅胶颗粒上。QIAEX II可从盐、琼脂糖、聚丙烯酰胺、染料、蛋白质和核酸中纯化DNA,无需苯酚提取或乙醇沉淀。QIAEX II对TAE和TBE缓冲液中的多种琼脂糖都有效。

QIAEX II颗粒确保高效回收DNA,且不被剪切,大片段的DNA也不例外。碘化钠是一种不容易从DNA样本中去除,还会影响后续反应的物质,优化的缓冲液可使回收的DNA中不含碘化钠。

QIAEX II体系使用的溶解和结合缓冲液包含一种特殊的pH指示剂。简单的颜色变化即可指示结合混合液的最佳pH值,确保DNA有效吸附到QIAEX II硅胶颗粒(参见"pH indicator dye")。有色染料也容易辨别结合混合液中未溶解的琼脂糖,确保完全溶解,获得最大的DNA产量。

Procedure

QIAEX II硅胶颗粒加入到溶解的凝胶碎片中,颗粒通过简单的离心操作收集(参见"QIAEX II procedure")。洗涤后,纯DNA用20 µl Tris缓冲液或水洗脱。

QIAEX II Gel Extraction Kit包括QIAEX II Suspension,结合和洗脱缓冲液,以及一本全面的手册。QIAEX II Suspension也可单独购买。有从琼脂糖凝胶、溶液和聚丙烯酰胺凝胶中纯化DNA的实验方案。

Applications

QIAEX II体系纯化得到的DNA可即用于多种应用,包括:

限制性酶切
标记
连接
PCR
Feature
Specifications
Binding capacity 5 µg/10 µl
Elution volume 20 µl
Format Tube
Fragment size 40 bp – 50 kb
Processing Manual
Recovery: oligonucleotides dsDNA Recovery: dsDNA fragments
Removal <10mers 17–40mers dye terminator proteins Removal <40mers
Sample type: applications DNA: PCR reactions
Technology Silica technology

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