MinElute Gel Extraction Kit

低溶出量で最高5 µgまでのDNAフラグメント(70 bp~4 kb)をゲルから抽出

S_1342_DNA_ME0803

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MinElute Gel Extraction Kit (50)

Cat. No. / ID:  28604

50 MinElute Spin Columns, Buffers, Collection Tubes (2 mL)
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¥18,000
Preparations
50
250

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 最小限の溶出量
  • 迅速な操作と容易な取り扱い
  • 再現性のある高い回収率
  • ゲル解析をより簡便化するGelPilot Loading Dye付き

Product Details

MinElute Gel Extraction Kitは、スピンカラム、バッファー、コレクションチューブにより構成され、シリカメンブレンによりDNAフラグメント(70 bp~4 kb)を400 mgまでのゲル切片から精製します。スピンカラムは微量(わずか10 µl)で溶出できるようにデザインされているため、高濃度のDNAが高収量で得られます。pH指示薬入りバッファーにより、DNAがスピンカラムに結合する最適なpHを簡単にチェックできます。MinEluteシステムで精製したDNAフラグメントはシークエンシング、マイクロアレイ解析、ライゲーション、トランスフォーメーション、制限酵素解析、標識反応、マイクロインジェクション、PCR、in vitro転写反応のような幅広いアプリケーションに即使用できます。

Performance

MinElute Gel Extraction Kit には、ヌクレオチド、酵素、塩類、アガロース、臭化エチジウムなどの夾雑物をDNAサンプルから除去するため、様々なダウンストリームアプリケーションに適した高濃度なDNAを提供します(図"高濃度なDNA精製")。

MinElute QIAquick Gel Extraction Kitは、ゲル抽出用のスピンカラムが含まれています。マイクロ遠心機または吸引マニホールドを使用することにより、70 bp~4 kbの高濃度なDNAが迅速に精製されます(4~10kbのDNAフラグメントの精製にはQIAquick Gel Extraction Kitを、70 bp以下あるいは10 kb以上のフラグメントにはQIAEX II Gel Extraction System をご利用ください)。

Principle

MinElute PCR Purification Kitでは、高塩濃度バッファーによりDNAを結合し、低塩濃度バッファーあるいは水によりDNAを溶出するシリカゲルメンブレン を利用しています。シリカメンブレンテクノロジーにより樹脂漏れ、および懸濁液関連の問題や不便さが解消されます。それぞれの用途に応じて至適化された結合バッファーにより、種々のサイズのDNAフラグメントを選択的に吸着します。

Gel Loading Dye

より迅速で簡便なサンプル処理と解析を実現するため、ゲル電気泳動用のローディング色素が付いています。GelPilot Loading Dye は3 種類のマーカー色素(xylene cyanol、bromophenol blueおよびorange G)が入っており、短いDNAフラグメントのゲルからの流出を回避でき、アガロースゲルの泳動時間の至適化が簡単に行なえます(図 " GelPilot Loading Dye")。

See figures

Procedure

MinEluteシステムでは結合-洗浄-溶出という簡単な操作だけでDNAのクリーンアップが可能です(フローチャート" MinElute操作手順"参照)。ゲル切片をpH指示薬を含んだバッファーに溶解させると、DNA結合の至適pHを容易に決めることができます(図  "pH 指示薬")。DNAを含むバッファーをMinElute Spin Column にアプライします。DNAは添付のバッファー中の高塩濃度の条件下でシリカゲルメンブレンに吸着します。不純物は洗い流され、純粋なDNAを添付の低塩濃度溶出バッファー、あるいは水で溶出します。得られたDNAは、その後のアプリケーションにも即使用可能です。

操作法

MinElute Spin Column は2つの簡便な操作法オプションのためにデザインされました(図" MinElute操作手順")。スピンカラムは簡便な卓上型マイクロ遠心機あるいはQIAvac 24 Plus のようなルアーコネクター付き吸引装置で使用できます。MinElute Gel Extraction Kitは他のQIAGENスピンカラムを利用したキット同様にQIAcubeでの完全自動化が可能で、 標準化された再現性の高い結果が得られます(図 "Spin Column操作オプション A B C D、およびE")。

See figures

Applications

MinEluteあるいはQIAquickシステムで精製したDNAフラグメントは以下のようなアプリケーションに即使用可能です。

  • 次世代シークエンシングを含むシークエンシング
  • マイクロアレイ解析
  • ライゲーションやトランスフォーメーション
  • 制限酵素分解
  • 標識反応

Supporting data and figures

Resources

MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
クイックスタートプロトコール (1)
Certificates of Analysis (1)
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