MinElute Gel Extraction Kit

用于在较低的洗脱体积下对多达 5 µg 的 DNA 片段(70 bp 至 4 kb)进行凝胶回收

S_1342_DNA_ME0803

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MinElute Gel Extraction Kit (50)

Cat. No. / ID:   28604

50 个 MinElute Spin Columns、Buffers、Collection Tubes (2 mL)
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Preparations
50
250
MinElute Gel Extraction Kit 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。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 极小的洗脱体积
  • 程序快速,操作简便
  • 回收率高,重复性好
  • 凝胶加载染料方便样本分析

产品详情

MinElute Gel Extraction Kit 提供离心柱、缓冲液和采样管,适用于从多达 400 mg 的凝胶切片中进行基于硅胶膜技术的 70 bp–4 kb 的 DNA 片段的纯化。离心柱的设计允许以非常小的体积(小至 10 µL)进行洗脱,获得高产量的高度浓缩的 DNA。集成的 pH 指示剂可轻松确定 DNA 吸附到离心柱的最佳 pH 值。用 MinElute 系统纯化的 DNA 片段可直接用于所有应用,包括测序、微阵列分析、连接和转化、限制性酶切、标记、微量注射、PCR 和体外转录。MinElute Gel Extraction Kit 可在 QIAcube Connect 上自动操作。

为获得最佳效果,建议将本产品与 QIAvac 24 Plus 一起使用。

绩效

MinElute Gel Extraction Kit 可去除 DNA 样本中的核苷酸、酶、盐、琼脂糖、溴化乙锭和其他杂质,提供适合各种下游应用的高浓度 DNA(见图“ 更高的 DNA 浓度”)。

MinElute Gel Extraction Kit 提供用于凝胶提取的离心柱。使用微型离心机或真空歧管,可快速纯化 70 bp –4 kb 的高浓度 DNA。(4 kb – 10 kb 的 DNA 片段应使用 QIAquick Gel Extraction Kit 纯化,小于 70 bp 或大于 10 kb 的 DNA 片段应使用 QIAEX II Gel Extraction System 提取)。

查看图表

原理

MinElute Kit 包含一个硅胶膜组件,用于在高盐缓冲液中结合 DNA,然后用低盐缓冲液或水进行洗脱。硅胶膜技术消除了树脂松散和稀浆带来的问题和不便。专用结合缓冲液针对特定应用进行了优化,可促进特定大小范围内 DNA 分子的选择性吸附。

凝胶加载染料

为了更快、更方便地进行样本处理和分析,我们提供了凝胶加载染料。GelPilot Loading Dye 包含三种跟踪染料(二甲苯胍青、溴酚蓝和橙黄 G),便于优化琼脂糖凝胶的运行时间,防止较小的 DNA 片段迁移过远(见图“ GelPilot Loading Dye”)。

查看图表

程序

MinElute 系统采用简单的结合-洗涤-洗脱程序(见流程图 “ MinElute 程序”)。将凝胶切片溶解在含有 pH 指示剂的缓冲液中,这样就能轻松确定 DNA 结合的最佳 pH 值(见图“ pH 指示基团染料”),然后将混合物加到 MinElute 离心柱上。核酸在缓冲液提供的高盐条件下吸附到硅胶膜上。杂质被洗去,用少量低盐缓冲液或水洗脱出的纯 DNA 可直接用于后续应用。

处理

MinElute 离心柱的设计提供了两种方便的处理选项。离心柱可安装到传统的台式微型离心机或任何带鲁尔接头的真空歧管上,如带有 QIAvac Luer Adapter 的 QIAvac 24 Plus。MinElute Gel Extraction Kit 以及其他基于 QIAGEN 离心柱的试剂盒均可在 QIAcube Connect 上实现全自动操作,从而提高生产效率和结果的规范化(见图“离心柱处理选项  A B C D 和  QIAcube Connect”)。

查看图表

应用

用 MinElute 或 QIAquick 系统纯化的 DNA 片段可直接用于所有应用,包括

  • 测序,包括新一代测序
  • 微阵列分析
  • 连接和转化
  • 限制性酶切
  • 标记

辅助数据和图表

Specifications

FeaturesSpecifications
Binding capacity5 µg
Elution volume10 µL
Fragment size70 bp – 4 kb
Sample type: applicationsDNA:PCR 反应
Technology凝胶提取
Recovery: oligonucleotides dsDNA回收率:dsDNA 片段
Format试管
Processing手动

资源

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
快速启动实验方案 (1)
试剂盒操作手册 (1)
MinElute Handbook
PDF (611KB)
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (1)
MinElute Handbook
PDF (611KB)
Quick-Start Protocols (1)

Publications

MicroRNA-137 targets microphthalmia-associated transcription factor in melanoma cell lines.
Bemis LT; Chen R; Amato CM; Classen EH; Robinson SE; Coffey DG; Erickson PF; Shellman YG; Robinson WA;
Cancer Res; 2008; 68 (5):1362-8 2008 Mar 1 PMID:18316599
Molecular and phylogenetic analyses reveal mammalian-like clockwork in the honey bee (Apis mellifera) and shed new light on the molecular evolution of the circadian clock.
Rubin EB; Shemesh Y; Cohen M; Elgavish S; Robertson HM; Bloch G;
Genome Res; 2006; 16 (11):1352-65 2006 Oct 25 PMID:17065608
An accurate fluorescent assay for quantifying the extent of RNA editing.
Roberson LM; Rosenthal JJ;
RNA; 2006; 12 (10):1907-12 2006 Sep 6 PMID:16957279
Adaptive evolution of fertilization proteins within a genus: variation in ZP2 and ZP3 in deer mice (Peromyscus).
Turner LM; Hoekstra HE;
Mol Biol Evol; 2006; 23 (9):1656-69 2006 Jun 14 PMID:16774977
Collision events between RNA polymerases in convergent transcription studied by atomic force microscopy.
Crampton N; Bonass WA; Kirkham J; Rivetti C; Thomson NH;
Nucleic Acids Res; 2006; 34 (19):5416-25 2006 Sep 29 PMID:17012275

FAQ

I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560
Can I store agarose gel slices containing DNA for gel extraction at a later point?
Cut out the slice of agarose containing the DNA fragment of interest, and store it at 4oC in an Eppendorf tube sealed with Parafilm.
FAQ ID -313
Are the columns of the MinElute Reaction Cleanup-, Gel Extraction-, and PCR Purification Kit identical?
Yes, and therefore they are interchangeable.
FAQ ID -581
Can a QIAquick Gel Extraction Kit be used to obtain RNA from a formaldehyde gel?

Yes. The QIAquick Gel Extraction Kit for extraction of DNA from gels can also be used for RNA gel extraction. Please see a user-developed procedure below, which was kindly provided by J. Knobloch, Heinrich Heine University, Düsseldorf, Germany. Note that this protocol has not been thoroughly tested and optimized by QIAGEN. QIAquick Gel Extraction Kits are not guaranteed to be RNase-free.

  1. Excise the RNA fragment from the formaldehyde agarose gel with a clean, sharp scalpel.
  2. Weigh the gel slice, and record the weight. Soak the gel slice in TE buffer for 25 min at room temperature with gentle shaking.
  3. Remove the gel slice from the TE buffer, and place it in a colorless tube. Add 6 volumes of Buffer QG to 1 volume of gel, based on the gel weight (100 mg ~ 100 µl).
  4. Incubate at 58°C for 25 min. To help dissolve the gel, mix by vortexing the tube every 2–3 min during the incubation.
  5. Continue the QIAquick Gel Extraction Kit Protocol (using a microcentrifuge) in the QIAquick Spin Handbook, beginning with step 4.
Please click here  for Figure 1. 

Figure 1:

A. Total RNA was isolated from the parasitic blood fluke Schistosoma mansoni using the RNeasy Mini Kit and run on a formaldehyde agarose (1.2%) gel. (Note: The 28S rRNA in S. mansoni contains a break site so that the rRNA splits into two parts, which run on a gel at the same size as the 18S rRNA.) The rRNA bands were excised and treated as described above (left lane) or using 10 volumes of Buffer QG in step 3 (right lane).

B. The extracted RNA was then analyzed on a new formaldehyde agarose gel. (Data kindly provided by J. Knobloch, Department of Genetic Parasitology, Heinrich Heine University, Düsseldorf, Germany).

FAQ ID -133
Why does my DNA sample float out of the slot when loading it onto an agarose gel?

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
Do you have information about the cleanup of single-stranded DNA (ssDNA) with QIAquick columns?

As a rule of thumb, single-stranded DNA binds to silica with approximately half the affinity of a double-stranded DNA fragment of the same length under the buffer conditions used in the QIAquick and MinElute Kits. Even though no systematic experimental data exists, we expect that recovery of ssDNA fragments of approximately 200 nucleotides and below will not be very efficient after cleanup using the QIAquick PCR Purification Kit or MinElute PCR Purification Kit. By comparison, it should be possible to purify fragments longer than 140 nucleotides using the QIAquick Gel Extraction Kit.

Note that recovery of single strand DNA is influenced to some degree also by factors such as base composition and secondary structure. It has to be determined empirically by the researcher if cleanup of single-stranded DNA with QIAquick columns yields satisfactory results.

FAQ ID -759
What is the composition of Buffer EB?

The composition of Buffer EB is:

  • 10 mM Tris-Cl, pH 8.5

Buffer EB is the elution buffer used in the QIAquick PCR, Gel Extraction, Nucleotide Removal Kits, and MinElute Kits for DNA cleanup, and the QIAprep Miniprep Kits for small-scale plasmid purification. The purified DNA can also be eluted in TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0), but the EDTA may inhibit subsequent enzymatic reactions.

FAQ ID -199
What is the small band below my fragment of interest on an agarose gel after DNA cleanup using QIAquick?

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
Can I buy QIAquick and MinElute columns separately?

The QIAquick Spin Columns (100) (cat. no. 28115) in the QIAquick PCR Purification, Gel Extraction, Nucleotide Removal and PCR & Gel Cleanup kits are also sold separately from the kits.

The MinElute columns in the MinElute PCR Purification, Gel Extraction and Reaction Cleanup kits are not sold separately.

We always provide extra buffers in our kits so you can scale up reactions, add extra washes or allow for spillage.

FAQ ID -2460
How can I extract DNA from a polyacrylamide (PAGE) gel?

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