QIAEX II 系统

用于从凝胶和溶液中纯化 DNA 片段（40 bp 至 50 kb）

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QIAEX II Gel Extraction Kit (150)

Cat. No. / ID: 20021

用于 150 次提取：3 x 0.5 ml QIAEX II 悬浮液、缓冲液

Reactions

150

500

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QIAEX II 系统旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

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✓ 博学专业的产品和技术支持

✓ 快速可靠的（再）订购

特点

有效提取 40 bp 至 50 kb 的 DNA
从 TAE 或 TBE 琼脂糖凝胶和聚丙烯酰胺凝胶中进行凝胶提取
无碘化钠干扰后续反应
不会剪切大的 DNA 片段

产品详情

QIAEX II 系统提供了一种二氧化硅颗粒悬浮液，在存在离液盐的情况下，DNA 片段与之结合。将 QIAEX II 悬浮液加入溶液或溶解的琼脂糖凝胶切片中，使其与 DNA 结合。通过短暂离心收集颗粒，进行清洗，然后用 Tris 缓冲液或水洗脱 40 bp 至 50 kb 的 DNA。

绩效

使用 QIAEX II 系统可有效回收 10 ng 至 10µg DNA（见图 “稳定的回收率”）。使用 30µl QIAEX II 悬浮液，可轻松将用于批量纯化凝胶片段的通用程序的结合能力放大到 15µg。

QIAEX II 系统提供的硅胶颗粒可纯化 60–95% 的 DNA 片段 (40 bp – 50 kb)。体积为 10µl 的 QIAEX II 悬浮液可结合多达 5µg 的 DNA，随后用 20µl 洗脱。

根据大小进行回收

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

* 将 2 µg 加载到 1% TAE 琼脂糖凝胶上。

查看图表

稳定的回收率。

原理

使用 QIAEX II 系统纯化 DNA 片段的基础是琼脂糖的溶解和核酸在离液盐存在的情况下对 QIAEX II 硅胶颗粒的选择性吸附。QIAEX II 能从盐类、琼脂糖、聚丙烯酰胺、染料、蛋白质和核苷酸中分离出 DNA，且无需苯酚提取或乙醇沉淀。QIAEX II 对 TAE 或 TBE 缓冲液中任何类型的琼脂糖都有效。

QIAEX II 颗粒可为凝胶提取提供稀浆，确保即使是大的 DNA 片段也能在不剪切的情况下高效回收。经过优化的缓冲液可以在不使用碘化钠的情况下回收 DNA，而碘化钠是很难从 DNA 样本中去除的，并且还可能会影响后续反应。

QIAEX II 系统使用的溶解和结合缓冲液含有独特的 pH 指示剂。简单的颜色变化就能显示结合混合物的 pH 值是否最适合 QIAEX II 硅胶颗粒对 DNA 的有效吸附（见图 “pH 指示基团染料”）。有颜色的染料还能让人轻松看到结合混合物中任何未溶解的琼脂糖，确保完全溶解以获得最大产量。

借助溶解和结合缓冲液中的 pH 指示基团染料，可以方便地通过目测确定 DNA 吸附的最佳 pH 值 (pH ≤7.5)。如果琼脂糖凝胶电泳缓冲液使用频繁或配制不正确，会导致结合混合物 pH 值不正确。在这种情况下，只需加入 10 µl 3 M 乙酸钠（pH 值为 5.0）即可轻松调节 pH 值。

查看图表

pH 指示基团染料。

程序

将 QIAEX II 硅胶颗粒加入已溶解的凝胶切片中，通过短暂的离心步骤收集颗粒（见流程图 “QIAEX II 程序”）。洗涤后，用 20 µl Tris 缓冲液或水洗脱纯 DNA 片段。

QIAEX II 系统提供 QIAEX II 悬浮液、结合缓冲液、洗涤缓冲液和一本全面的手册。提供了从琼脂糖凝胶、溶液和聚丙烯酰胺凝胶中纯化 DNA 的方案。

查看图表

QIAEX II 程序。

应用

用 QIAEX II 系统纯化的 DNA 可直接用于大多数应用，包括

限制性酶切
标记
连接
PCR

特点	规格
结合能力	5 µg/10 µl
洗脱体积	20 µl
格式	试管
片段大小	40 bp – 50 kb
处理	手动
回收率：寡核苷酸 dsDNA	回收率：dsDNA 片段
去除 <10mer 17–40mer 染料终止物蛋白质	去除 <40mer
样本类型：应用	DNA：PCR 反应
技术	硅胶膜技术

辅助数据和图表

稳定的回收率。

图为使用 QIAEX II Gel Extraction Kit 从 1% 琼脂糖凝胶中提取的不同量的 2.9 kb DNA 片段。

资源

Download Safety Data Sheets for QIAGEN product components.

Publications

T-B+NK+ severe combined immunodeficiency caused by complete deficiency of the CD3zeta subunit of the T-cell antigen receptor complex.

Roberts JL; Lauritsen JP; Cooney M; Parrott RE; Sajaroff EO; Win CM; Keller MD; Carpenter JH; Carabana J; Krangel MS; Sarzotti M; Zhong XP; Wiest DL; Buckley RH;

Blood; 2006; 109 (8):3198-206 2006 Dec 14 PMID:17170122

Pharmacologic inhibition of epigenetic modifications, coupled with gene expression profiling, reveals novel targets of aberrant DNA methylation and histone deacetylation in lung cancer.

Zhong S; Fields CR; Su N; Pan YX; Robertson KD;

Oncogene; 2006; 26 (18):2621-34 2006 Oct 9 PMID:17043644

Role for nonstructural protein 1 of severe acute respiratory syndrome coronavirus in chemokine dysregulation.

Law AH; Lee DC; Cheung BK; Yim HC; Lau AS;

J Virol; 2006; 81 (1):416-22 2006 Oct 11 PMID:17035307

Effects of the chemotherapeutic agent doxorubicin on the protein C anticoagulant pathway.

Woodley-Cook J; Shin LY; Swystun L; Caruso S; Beaudin S; Liaw PC;

Mol Cancer Ther; 2006; 5 (12):3303-11 2006 Dec PMID:17172434

Exportin-5 orthologues are functionally divergent among species.

Shibata S; Sasaki M; Miki T; Shimamoto A; Furuichi Y; Katahira J; Yoneda Y;

Nucleic Acids Res; 2006; 34 (17):4711-21 2006 Sep 8 PMID:16963774

FAQ

Larger DNA fragments bind more tightly to the QIAquick columns. It is difficult to predict whether a DNA fragment larger than 10 kb can be efficiently recovered, because this depends on base composition as well as fragment size. If the fragment is only a few kb larger than the 10 kb limit, it can be helpful to heat the elution buffer EB to 60°C and let it incubate on the column for a few minutes before centrifuging. However, please note that it will become less likely to recover your sample the larger the fragment size is. As we cannot guarantee recovery of fragments larger than the maximum cutoff size, we do not recommend to purify such fragments using QIAquick Kits.

The QIAEX II Kit can be used to extract DNA fragments up to 50 kb from agarose or polyacrylamide gels.

FAQ ID -756

Assuming that the ligation conditions are correct, QIAEX II particle carryover may affect ligation efficiency. Under low salt conditions, enzymes may bind to QIAEX II particles, thus reducing enzymatic activity in the ligation reaction. To remove the particles, centrifuge the tube containing the DNA for 30 seconds before pipetting the DNA.

FAQ ID -141

Cut out the slice of agarose containing the DNA fragment of interest, and store it at 4^oC in an Eppendorf tube sealed with Parafilm.

FAQ ID -313

Yes, single stranded DNA can be isolated from agarose or polyacrylamide gels using the QIAEX II Gel Extraction Kit.

FAQ ID -576

DNA fragments purified with the QIAGEN DNA Cleanup Systems, i.e., the QIAquick PCR Purification Kit, the MinElute Reaction Cleanup Kit, the QIAEX II Gel Extraction Kit etc. may float out of the loading wells of agarose gels due to residual ethanol carried over from the wash step with Buffer PE (despite the addtition of glycerol-containing loading buffer).

Use either of the following options to remove residual ethanol from the eluate:

re-purify the sample using a QIAquick-, or MinElute column, or QIAEX II resin
incubate the eluate at 56°C for 10 min to evaporate the ethanol
dry down the sample in a vacuum centrifuge, and resuspend the pellet in a small volume of sterile water

FAQ ID -205

Yes, bisulfite containing methylation reactions can be cleaned up with our silica-based cleanup products, such as QIAquick and QIAEX II. Please see Goyon et al. (1994), 'Perpetuation of cytosine methylation in Ascobolus immersus implies a novel type of maintenance methylase', published in J Mol Biol. 1994 Jul 1;240(1):42-51, for a reference.

FAQ ID -519

Occasionally, DNA fragments eluted from the silica matrix of QIAquick, MinElute or QIAEX II Kits will contain denatured single-stranded DNA (ssDNA), appearing as a smaller band on an analytical gel. Under certain conditions, chaotropic agents (present in all silica-based DNA purification methods) can denature DNA fragments. This is a rare event that may be influenced by sequence characteristics such as the presence of inverted repeats or A–T-rich stretches.

Because salt and buffering agents promote renaturation of DNA strands, the following tips are recommended:

use the eluted DNA to prepare your downstream enzymatic reaction, but omit the enzyme. Incubate the reaction mix at 95°C for 2 minutes to reanneal the ssDNA, and allow the tube to cool slowly to room temperature before adding the enzyme and proceeding
alternatively, the DNA can be eluted from the silica-gel membrane or resin in 10 mM Tris buffer containing 10 mM NaCl. However, the salt concentration of the eluate must then be taken into consideration in downstream applications.

FAQ ID -148

The QIAEX II and QIAquick Gel Extraction Kit can be used to extract DNA from polyacrylamide gels.

The QIAEX II Handbook contains a protocol for Polyacrylamide Gel Extraction. A specialized User-Developed Protocol (QQ05) is available when using the QIAquick Gel Extraction Kit for this purpose.

Both protocols require the preparation of a diffusion buffer and a disposable plastic column or syringe barrel containing a Whatman GF/C filter or siliconized glass wool. To ensure optimal diffusion, cut the gel slices as small as possible, and use 2 volumes of diffusion buffer per 1 volume of gel. Increasing incubation time (protocol step 3) may result in higher yields.

FAQ ID -120