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MinElute Gel Extraction Kit

적은 용출량에서 최대 5µg DNA 절편(70bp~4kb)의 겔 추출에 사용합니다

S_1342_DNA_ME0803

✓ 연중무휴 하루 24시간 자동 온라인 주문 처리

✓ 풍부한 지식과 전문성을 갖춘 제품 및 기술 지원

✓ 신속하고 안정적인 (재)주문

MinElute Gel Extraction Kit (50)

Cat. No. / ID:   28604

MinElute 스핀 컬럼 50개, 완충액, Collection Tubes (2 mL)
Preparations
50
250
MinElute Gel Extraction Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.

✓ 연중무휴 하루 24시간 자동 온라인 주문 처리

✓ 풍부한 지식과 전문성을 갖춘 제품 및 기술 지원

✓ 신속하고 안정적인 (재)주문

특징

  • 매우 적은 용출량
  • 신속한 절차 및 간편한 취급 방법
  • 재현성이 뛰어난 높은 회수율
  • 편리한 샘플 분석을 위한 겔 로딩 염료

제품 세부 정보

MinElute Gel Extraction Kit는 최대 400mg의 겔 절편에서 70bp~4kb의 DNA 절편을 실리카 막 기반으로 정제할 수 있는 스핀 컬럼, 완충액, collection 튜브를 제공합니다. 스핀 컬럼은 매우 적은 용량(최소 10µl)으로 용출할 수 있도록 설계되어 고농축 DNA를 높은 수율로 추출할 수 있습니다. 포함된 pH indicator 염료를 통해 스핀 컬럼에 결합하는 DNA에 대한 최적의 pH를 쉽게 파악할 수 있습니다. MinElute system으로 정제된 DNA 절편은 염기서열 분석, 마이크로어레이 분석, 결찰(ligation) 및 형질전환, 제한효소 처리(restriction digestion), 라벨링, 미세 주입, PCR 및 체외 전사를 포함한 모든 응용 분야에서 바로 사용할 수 있습니다. MinElute Gel Extraction Kit는 QIAcube Connect에서 자동화할 수 있습니다.

최적의 결과를 얻으려면 이 제품을 QIAvac 24 Plus와 함께 사용하는 것이 좋습니다.

성능

MinElute Gel Extraction Kit는 DNA 샘플에서 뉴클레오타이드, 효소, 염, 아가로스, 브로민화 에티듐(ethidium bromide) 및 기타 불순물을 제거하여 다양한 다운스트림 응용 분야에 적합한 고농축 DNA를 제공합니다(그림 " 고농축 DNA" 참조).

MinElute Gel Extraction Kit는 겔 추출을 위한 스핀 컬럼을 제공합니다. 마이크로 원심분리기 또는 진공 매니폴드를 사용하여 70bp~4kb의 고농도 DNA를 빠르게 정제합니다. (4kb~10kb의 DNA 절편은 QIAquick Gel Extraction Kit를 사용하여 정제해야 하며, 70bp보다 작거나 10kb보다 큰 DNA 절편은 QIAEX II Gel Extraction System으로 추출해야 합니다).

그림 참조

원리

MinElute Kit에는 염도가 높은 완충액에서 DNA를 결합하고 저염 완충액 또는 물로 용출하기 위한 실리카 막 어셈블리가 포함되어 있습니다. 실리카 막 기술은 분산 수지 및 슬러리(slurry)와 관련된 문제와 불편을 해소합니다. 특수 결합 완충액은 각 응용 분야에 최적화되어 있으며 특정 크기 범위 내에서 DNA 분자를 선택적으로 흡착하도록 합니다.

겔 로딩 염료

더 빠르고 편리한 샘플 처리 및 분석을 위해 겔 로딩 염료가 제공됩니다. GelPilot 로딩 염료는 아가로스 겔 실행 시간을 최적화하고 작은 DNA 절편이 너무 멀리 이동하는 것을 방지하기 위해 세 가지 추적 염료(크실렌 시아놀, 브로모페놀 블루, 오렌지 G)를 함유하고 있습니다(그림 " GelPilot 로딩 염료" 참조).

그림 참조

절차

MinElute system은 간단한 결합-세척-용출 절차를 사용합니다(순서도 " MinElute 절차" 참조). 겔 절편을 pH indicator 염료가 포함된 완충액에 용해하여 DNA 결합을 위한 최적의 pH를 쉽게 확인할 수 있으며(그림 " pH indicator 염료" 참조), 혼합물을 MinElute pin column에 적용합니다. 핵산은 완충액의 높은 염도 조건에서 실리카겔 막에 흡착됩니다. 불순물을 씻어내고 제공된 소량의 저염 완충액 또는 물로 순수한 DNA를 용출하여 후속 응용 분야에 바로 사용할 수 있습니다.

취급

MinElute 스핀 컬럼은 두 가지 편리한 취급 옵션을 제공하도록 고안되었습니다. 스핀 컬럼은 기존의 탁상용 마이크로 원심분리기 또는 루어 커넥터가 있는 모든 진공 매니폴드(예: QIAvac Luer Adapters가 있는 QIAvac 24 Plus)에 장착할 수 있습니다. MinElute Gel Extraction Kit는 다른 QIAGEN 스핀 컬럼 기반 키트와 더불어 QIAcube Connect에서 완전히 자동화할 수 있어 생산성을 높이고 결과를 표준화할 수 있습니다 (그림 "스핀 컬럼 취급 옵션  A,  B,  C,  D 및  QIAcube Connect" 참조).

그림 참조

응용 분야

MinElute 또는 QIAquick System으로 정제된 DNA 절편은 다음을 포함한 모든 응용 분야에서 바로 사용할 수 있습니다.

  • 차세대 염기서열 분석을 포함한 염기서열 분석
  • 마이크로어레이 분석
  • 결찰(ligation) 및 형질전환
  • 제한효소 처리(Restriction digestion)
  • 라벨링

지원되는 데이터 및 수치

Specifications

FeaturesSpecifications
Binding capacity5µg
Elution volume10µL
Fragment size70bp~4kb
Sample type: applicationsDNA: PCR 반응
Technology겔 추출
Recovery: oligonucleotides dsDNA회수: dsDNA 절편
Format튜브
Processing수동

리소스

Safety Data Sheets (1)
Quick-Start Protocols (1)
Kit Handbooks (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
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