QIAquick Gel Extraction Kit

겔 또는 효소 반응에서 최대 10µg DNA(70bp~10kb)의 겔 추출/클린업에 사용합니다

S_1341_DNA_QQ0799

✓ 연중무휴 하루 24시간 자동 온라인 주문 처리

✓ 풍부한 지식과 전문성을 갖춘 제품 및 기술 지원

✓ 신속하고 안정적인 (재)주문

QIAquick Gel Extraction Kit (50)

Cat. No. / ID:   28704

50회 반응분 겔 추출 또는 클린업용: QIAquick 스핀 컬럼 50개, 완충액, Collection Tubes (2 ml)
₩159,000.00
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KitColumn
QIAquick Gel Extraction Kit
QIAquick PCR & Gel Cleanup Kit
QIAquick Spin Columns
Reactions
50
250
1000
QIAquick 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

  • 바로 사용 가능한 DNA 최대 95% 회수
  • 빠르고 편리한 절차
  • 간단한 3단계로 최대 10kb의 DNA 클린업
  • 편리한 샘플 분석을 위한 겔 로딩 염료
  • 겔 추출 및 PCR 클린업을 위한 결합 키트

제품 세부 정보

QIAquick Gel Extraction Kit는 겔(최대 400mg 절편) 또는 효소 반응에서 DNA 절편의 실리카 막 기반 정제를 위한 스핀 컬럼, 완충액, collection 튜브를 제공합니다. 간단하고 빠른 결합-세척-용출 절차 및 30~50µl의 용출량으로 70bp~10kb 범위의 DNA를 정제합니다. 포함된 pH indicator 염료를 통해 스핀 컬럼에 결합하는 DNA에 대한 최적의 pH를 쉽게 파악할 수 있습니다. 또한 QIAquick PCR & Gel Cleanup Kit는 >100bp의 PCR 산물과 최대 10kb의 DNA를 정제하기 위한 완충액을 제공합니다. 해당 절차는 QIAcube Connect에서 완전히 자동화할 수 있습니다.

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

성능

QIAquick Gel Extraction Kit를 사용하면 샘플에서 뉴클레오타이드, 효소, 염, 아가로스, 브로민화 에티듐(ethidium bromide) 및 기타 불순물을 제거하여 최대 80%의 DNA를 회수할 수 있습니다(그림 " 겔에서 높은 회수율" 참조). 마이크로 원심분리기 또는 진공 매니폴드를 사용하여 1~24개 샘플에서 70bp~10kb 범위의 DNA를 정제합니다. 정제된 DNA는 예를 들어 염기서열 분석에 사용할 수 있습니다(그림 " 겔 추출 후 신뢰할 수 있는 염기서열 분석" 참조). 70bp보다 작거나 10kb보다 큰 DNA 절편은 QIAEX II Gel Extraction System으로 추출해야 합니다.

QIAquick PCR Purification 절차는 DNA 샘플에서 프라이머, 뉴클레오타이드, 효소, 미네랄 오일, 염 및 기타 불순물을 제거합니다(그림 " PCR 후 프라이머 완전 제거" 참조). 마이크로 원심분리기 또는 진공 매니폴드를 사용하여 100bp~10kb 범위의 DNA를 정제합니다.

그림 참조

원리

QIAquick Kit에는 염도가 높은 완충액에서 DNA를 결합하고 저염 완충액 또는 물로 용출하기 위한 실리카 막 어셈블리가 포함되어 있습니다. 정제 절차는 DNA 샘플에서 프라이머, 뉴클레오타이드, 효소, 미네랄 오일, 염, 아가로스, 브로민화 에티듐(ethidium bromide) 및 기타 불순물을 제거합니다(그림 " 겔에서 높은 회수율" 참조). 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

QIAquick system은 간단한 결합-세척-용출 절차를 사용합니다(순서도 " QIAquick 및 MinElute 절차" 참조). 겔 절편을 DNA 결합을 위한 최적의 pH를 쉽게 확인할 수 있는 pH indicator 염료가 포함된 완충액에서 용해하고, 혼합물을 QIAquick 스핀 컬럼에 적용합니다(그림 " pH indicator 염료" 참조). 핵산은 완충액의 높은 염도 조건에서 실리카 막에 흡착됩니다. 불순물을 씻어내고 제공된 소량의 저염 완충액 또는 물로 순수한 DNA를 용출하여 모든 후속 응용 분야에 바로 사용할 수 있습니다.

취급

QIAquick 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. QIAquick 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

QIAquick system으로 정제된 DNA 절편은 염기서열 분석, 결찰(ligation) 및 형질전환, 제한효소 처리(restriction digestion), 라벨링, 미세 주입, PCR 및 체외 전사를 포함한 모든 응용 분야에서 바로 사용할 수 있습니다.

지원되는 데이터 및 수치

Specifications

FeaturesSpecifications
Binding capacity10µg
Format튜브
Fragment size70bp~10kb
Recovery: oligonucleotides dsDNA회수: dsDNA 절편
Processing수동
Removal <10mers 17–40mers dye terminator proteins<10mers 제거
Elution volume30~50µl
Technology실리카 기술
Sample type: applicationsDNA: PCR 반응

리소스

Kit Handbooks (1)
Safety Data Sheets (1)
Scientific Posters (1)
Certificates of Analysis (1)

Publications

STAT5 represses BCL6 expression by binding to a regulatory region frequently mutated in lymphomas.
Walker SR; Nelson EA; Frank DA;
Oncogene; 2006; 26 (2):224-33 2006 Jul 3 PMID:16819511

FAQ

I bound an 11 kb DNA fragment to a QIAquick column; is it completely lost?

Larger DNA fragments bind more tightly to the QIAquick columns. It is difficult to predict whether a DNA fragment larger than 10 kb can be efficiently recovered, because this depends on base composition as well as fragment size. If the fragment is only a few kb larger than the 10 kb limit, it can be helpful to heat the elution buffer EB to 60°C and let it incubate on the column for a few minutes before centrifuging. However, please note that it will become less likely to recover your sample the larger the fragment size is. As we cannot guarantee recovery of fragments larger than the maximum cutoff size, we do not recommend to purify such fragments using QIAquick Kits.

The QIAEX II Kit can be used to extract DNA fragments up to 50 kb from agarose or polyacrylamide gels.

 

FAQ ID -756
Are Buffer PB of the QIAquick PCR Purification Kit and Buffer QG of the QIAquick Gel Extraction Kit interchangeable?

Buffer PB of the QIAquick PCR Purification Kit cannot be used to extract DNA from agarose gels. However, Buffer QG of the QIAquick Gel Extraction Kit can be used to remove salt and proteins from enzymatic reactions by adding 3 volumes of Buffer QG and 1 volume of isopropanol to the reaction and proceeding with step 6 of the Gel Extraction Spin Protocol in the QIAquick Spin Handbook. See the QIAquick Spin Handbook for a list of reactions which can be cleaned up with the various QIAquick kits.

FAQ ID -786
Are QIAprep and QIAquick Spin columns interchangeable?
No. While columns from the QIAprep Spin Miniprep Kit and the QIAquick PCR Purification- and Gel Extraction Kits are based on silica-gel-membrane technology, each is designed to work optimally within its own kit format. In addition, the binding capacity and DNA recovery size cut-offs of the QIAprep and QIAquick Spin columns are different. QIAprep Spin columns bind up to 20 ug of plasmid DNA up to 50 kb in length, while the QIAquick Spin columns bind up to 10 ug of DNA with a maximum fragment size of 10 kb.
FAQ ID -311
Are the columns of the QIAquick PCR Purification-, Gel Extraction-, and Nucleotide Removal Kit interchangeable?
Yes - all QIAquick Spin Kits contain identical columns, but different binding buffers optimized for each specific application.
FAQ ID -577
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
How can I improve recoveries when using the QIAquick Kits?

Buffer PE did not contain ethanol

Ethanol must be added to Buffer PE (concentrate) before use. Repeat procedure with correctly prepared Buffer PE.

Inappropriate elution buffer

DNA will only be eluted efficiently in the presence of low-salt buffer (e.g., Buffer EB: 10 mM Tris·Cl, pH 8.5) or water. Elution efficiency is strongly dependent on the salt concentration and pH of the elution buffer. Contrary to adsorption, elution is most efficient under basic conditions and low salt concentrations. DNA is eluted with 50 or 30 µl of the provided Buffer EB (10 mM Tris·Cl, pH 8.5), or water. The maximum elution efficiency is achieved between pH 7.0 and 8.5. When using water to elute, make sure that the pH is within this range. In addition, DNA must be stored at –20°C when eluted with water since DNA may degrade in the absence of a buffering agent. Elution with TE (10 mM Tris·Cl, 1 mM EDTA, pH 8.0) is possible, but not recommended because EDTA may inhibit subsequent enzymatic reactions.

Elution buffer incorrectly dispensed

Add elution buffer to the center of the QIAquick membrane to ensure that the buffer completely covers the membrane. This is particularly important when using small elution volumes (30 µl).

FAQ ID -180
How do I perform a DNA precipitation to concentrate my sample?
  • Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
  • Mix, and store at –20°C for at least 1 h to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min
  • Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
  • Allow the DNA pellet to air-dry
  • resuspend the DNA in a suitable volume of sterile TE buffer or distilled water
FAQ ID -305
Will the QIAquick PCR Purification Kit remove sufficient SYBR Green from real-time PCR reactions to allow sequencing?

Based on our experience, the QIAquick PCR Purification Kit removes SYBR Green dye efficiently from PCR reactions.

We strongly assume that the QIAquick Gel Extraction Kit will be equally efficient in removing this dye, however, we recommend a 5 min incubation with wash Buffer PE on the QIAquick spin column at step 10 of the QIAquick Gel Extraction Kit Protocol. Efficiency of SYBR Green dye removal has to be validated by the enduser.

 

FAQ ID -637
Do you have protocols for multiple extractions of DNA fragments from agarose gels?

Yes, please follow the Supplementary Protocols 'High-throughput gel extractions using the QIAquick 96 PCR Purification Kit' (QQ03).  Please contact your local QIAGEN Technical Service for this protocol.

FAQ ID -944
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
Is it possible to clean up a methylation reaction containing bisulfite with QIAquick Cleanup Kits?
Yes, bisulfite containing methylation reactions can be cleaned up with our silica-based cleanup products, such as QIAquick and QIAEX II. Please see Goyon et al. (1994),  'Perpetuation of cytosine methylation in Ascobolus immersus implies a novel type of maintenance methylase', published in J Mol Biol. 1994 Jul 1;240(1):42-51, for a reference.
FAQ ID -519
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 do I safely inactivate biohazardous flow-through material?

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12
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