MinElute Gel Extraction Kit

少量の溶出で最大5 µg のDNAフラグメント（70 bp ～ 4 kb）のゲル抽出に

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

MinElute Gel Extraction Kit (50)

Cat. No. / ID: 28604

50 MinElute Spin Columns, Buffers, Collection Tubes (2 mL)

Preparations

250

Inquire

MinElute Gel Extraction Kitは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

特徴

非常に少量の溶出液量
迅速かつ簡単な操作
再現性と高い回収率
サンプル操作を便利にするローディングダイ

製品詳細

MinElute Gel Extraction Kitには、最大400 mgのゲルから70 bp – 4 kb DNAフラグメント精製にシリカメンブレンスピンカラム、バッファーおよびコレクションチューブが含まれます。スピンカラムは、非常に少量（10 µL）の溶出で濃縮DNAを高収率で得られるよう設計されています。pH指示薬により、スピンカラムへのDNA結合の至適pHを容易に確認できます。MinEluteで精製された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、ライゲーションと形質転換、制限酵素消化、標識、マイクロインジェクション、PCR、in vitro転写など様々なアプリケーションに直接使用できます。MinElute Gel Extraction Kitは、QIAcube Connectで自動化できます。

最適な結果を得るには、本製品をQIAvac 24 Plusと併用することをお勧めします。

パフォーマンス

MinElute Gel Extraction Kitは、DNAサンプルからヌクレオチド、酵素、塩、アガロース、臭化エチジウムなどの不純物を除去し、ダウンストリームのさまざまなアプリケーションへの高濃縮DNAが得られます（「高濃縮DNA」の図を参照）。

MinElute Gel Extraction Kitは、ゲル抽出用のスピンカラムキットです。小型遠心機または真空マニホールドを使用して、70 bp – 4 kbの高濃度の DNA を迅速に精製します。（4 kb – 10 kbのDNAフラグメントはQIAquick Gel Extraction Kitで、また70 bpより小さいまたは10 kbより大きいDNAフラグメントはQIAEX II Gel Extraction Systemで精製してください。）

図参照

より高濃度のDNA。

原理

MinElute Kitsは、高塩濃度バッファーでDNAを結合し、低塩濃度バッファーまたは水で溶出するためのシリカメンブレンキットです。シリカメンブレン技術は、緩い樹脂やスラリーによる問題や不都合がありません。特殊な結合バッファーは、特定のアプリケーションに最適化され、特定のサイズ内のDNA分子の選択的な吸着を促進します。

Gel loading dye

ローディングダイは、迅速で便利なサンプル処理と分析を可能にします。GelPilot Loading Dyeは、3種類のトラッキングダイ（キシレンシアノール、ブロモフェノールブルー、およびオレンジG）を含み、アガロールゲルのランタイムの最適化を容易にし、より小さなDNAフラグメントの過度な移動を防ぎます（「 GelPilot Loading Dye」の図を参照）。

図参照

GelPilot Loading Dye。

操作手順

MinEluteシステムは、シンプルな結合－洗浄－溶出の操作です（「 MinEluteの操作手順」のフローチャートを参照）。ゲルスライスは、pH指示薬を含むバッファーに溶解するので、DNA結合の至適pHを決定でき（「 pH Indicator Dye」の図を参照）、その混合液をMinEluteスピンカラムにアプライします。核酸は、バッファーによる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は、少量の低塩濃度バッファーまたは水と共に溶出し、後のアプリケーションに使用できます。

取り扱い

MinEluteスピンカラムは、2通りの操作方法が設定されてます。スピンカラムは、小型遠心機または QIAvac Luer Adapters付きのQIAvac 24 Plusなどのルアーコネクター付きの真空マニホールド上にフィットします。MinElute Gel Extraction Kitは、その他のQIAGENスピンカラムキットに加えて、QIAcube Connectで完全自動化して、生産性の向上、結果の標準化することができます（「スピンカラムの取り扱いオプション A、 B、 C、 D および QIAcube Connect」の図を参照）。

図参照

アプリケーション

MinEluteまたはQIAquick Systemで精製したDNAフラグメントは、以下を含むすべてのアプリケーションで使用できます。

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
ライゲーションと形質転換
制限酵素処理
Labeling

裏付けデータと数値

スピンカラム取り扱いオプション — C。

ルアーコネクター付きのマニホールド

Specifications

Features	Specifications
Binding capacity	5 µg
Elution volume	10 µl
Fragment size	70 bp ～ 4 kb
Sample type: applications	DNA：PCR反応
Technology	ゲル抽出
Recovery: oligonucleotides dsDNA	回収：dsDNAフラグメント
Format	チューブ
Processing	手動

リソース

Download Safety Data Sheets for QIAGEN product components.

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

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

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, and therefore they are interchangeable.

FAQ ID -581

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.

Excise the RNA fragment from the formaldehyde agarose gel with a clean, sharp scalpel.
Weigh the gel slice, and record the weight. Soak the gel slice in TE buffer for 25 min at room temperature with gentle shaking.
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).
Incubate at 58°C for 25 min. To help dissolve the gel, mix by vortexing the tube every 2–3 min during the incubation.
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

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

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

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

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 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

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