MinElute Gel Extraction Kit

低洗脱体积,从凝胶中纯化多达5 μg的DNA片段(70 bp到4 kb)

S_1342_DNA_ME0803

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

MinElute Gel Extraction Kit (50)

Cat. No. / ID:  28604

50 MinElute Spin Columns, Buffers, Collection Tubes (2 mL)
Copy order details
Log in To see your account pricing.
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。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 非常小的洗脱体积
  • 纯化速度快,操作简单
  • 回收率高,重复性好
  • 含有凝胶上样染料,方便样本分析

Product Details

MinElute Gel Extraction Kit含有离心柱、缓冲液和收集管,用于基于硅胶膜技术的DNA片段纯化,适用于从多达400 mg的凝胶胶条中纯化70 bp到4 kb的DNA。特殊设计的离心柱可将DNA洗脱至非常小的体积(10 μl),获得高产、高度浓缩的DNA。通过内置的pH指示剂可容易确定DNA结合到离心柱的最佳pH值。使用MinElute体系纯化的DNA片段可即用于各种应用,包括测序、微阵列分析、连接和转化、限制性酶切、标记、显微注射、PCR和体外转录。

Performance

MinElute Gel Extraction Kit纯化去除引物、核苷酸、酶、矿物油、盐、琼脂糖、溴化乙锭和DNA样品中的其他杂质,获得的高度浓缩的DNA可用于各种下游应用(参见" Higher DNA concentrations")。

MinElute QIAquick Gel Extraction Kit提供用于凝胶纯化的离心柱。使用微型离心机或真空装置快速纯化70 bp–4 kb的高度浓缩的DNA。(4 kb–10 kb的DNA片段使用QIAquick Gel Extraction Kit纯化,而小于70 bp或大于10 kb的片段使用QIAEX II Gel Extraction System纯化。)

See figures

Principle

MinElute Kits采用硅胶膜式纯化柱,在高盐条件下结合DNA,低盐或水可洗脱DNA。硅胶膜技术避免了松散树脂和悬液状态的问题及不方便性。经优化的特制结合缓冲液,专用于选择性吸附特定大小范围内的DNA分子。

凝胶上样染料

为更快速、更方便地进行分析,提供上样染料。GelPilot Loading Dye含有3种示踪染料(xylene cyanol、bromophenol blue和orange G),便于优化凝胶运行时间,避免小片段DNA跑得过远(参见" GelPilot Loading Dye")。

See figures

Procedure

MinElute体系应用简单的结合-洗涤-洗脱步骤(参见" MinElute procedure")。凝胶碎片溶解在含有pH指示剂的缓冲液中,可方便确定DNA结合的最佳pH值(参见 pH Indicator Dye),然后将混合物装载到MinElute离心柱上。在高盐条件下,核酸吸附在硅胶膜上。洗去杂质并用少量的低盐缓冲液或水洗脱DNA,可即用于各种下游应用。

操作

MinElute离心柱提供两种操作(参见" MinElute procedure")。将离心柱放入传统的微型离心机或任何含有连接器的真空装置上,如带有QIAvac Luer Adapters的QIAvac 24 Plus。MinElute Gel Extraction Kit和QIAGEN的其他离心柱试剂盒,可在QIAcube全自动核酸纯化仪上全自动进行,可提高产量,获得标准化结果。

See figures

Applications

MinElute或QIAquick System纯化的DNA片段可直接用于多种下游应用,包括:

  • 测序,包括第二代测序
  • 微阵列分析
  • 连接和转化
  • 限制性酶切
  • 标记

Supporting data and figures

Resources

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)
Certificates of Analysis (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