QIAquick PCR Purification Kit for PCR Cleanup

100bp~10kb의 최대 10µg PCR 산물 정제에 사용합니다

✓ 연중무휴 하루 24시간 자동 온라인 주문 처리

✓ 풍부한 지식과 전문성을 갖춘 제품 및 기술 지원

✓ 신속하고 안정적인 (재)주문

QIAquick PCR Purification Kit (50)

Cat. No. / ID:   28104

50회의 PCR 반응 정제용: QIAquick 스핀 컬럼 50개, 완충액, Collection Tubes (2 ml)
₩148,000.00
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KitColumn
QIAquick PCR Purification Kit
QIAquick PCR & Gel Cleanup Kit
QIAquick Spin Columns
Preparations
50
250
QIAquick PCR Purification 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
  • 간단한 3단계로 최대 10kb의 DNA 클린업
  • 편리한 샘플 분석을 위한 겔 로딩 염료
  • 겔 추출 및 PCR 클린업을 위한 결합 키트
  • 스핀 컬럼은 별도 구매 가능 

제품 세부 정보

QIAquick PCR Purification Kit는 >100bp PCR 산물의 실리카 막 기반 정제를 위한 스핀 컬럼, 완충액, collection 튜브를 제공합니다. 간단하고 빠른 결합-세척-용출 절차 및 30~50μl의 용출량으로 최대 10kb의 DNA를 정제합니다. 옵션으로 제공되는 pH indicator 염료를 통해 스핀 컬럼에 결합하는 DNA에 대한 최적의 pH를 쉽게 파악할 수 있습니다. 해당 절차는 QIAcube Connect에서 완전히 자동화할 수 있습니다.

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

QIAquick PCR Purification 표준 프로토콜은 또한 TRACKMAN Connected system을 이용하여 PIPETMAN M Connected 피펫과 함께(모두 Gilson 제품) 실행할 수 있습니다. TRACKMAN Connected system은 연구자에게 QIAquick PCR Purification 프로토콜을 안내하고 Bluetooth 지원 PIPETMAN M Connected 피펫 설정을 자동으로 조정합니다. 프로토콜 실행의 각 단계가 기록되어 종합적인 실행 보고서를 생성하므로 보고 속도를 높일 수 있습니다. 더 많은 정보 다운로드.

성능

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 " PCR 후 프라이머 완전 제거" 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). 결합 완충액을 PCR 샘플 또는 기타 효소 반응물에 직접 첨가하고 혼합물을 QIAquick 스핀 컬럼에 적용합니다. 결합 완충액에는 pH indicator 염료가 포함되어 있어 DNA 결합을 위한 최적의 pH를 쉽게 확인할 수 있습니다(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 PCR Purification Kit는 다른 QIAGEN 스핀 컬럼 기반 키트와 더불어 QIAcube에서 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, 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.

지원되는 데이터 및 수치

Specifications

FeaturesSpecifications
Binding capacity10µg
Technology실리카 기술
Sample type: applicationsPCR 및 기타 효소 반응에서 얻은 ssDNA 또는 dsDNA
Fragments removed< 40mers
Elution volume>30µl
Fragment size100bp~10kb
Processing수동
Format튜브
Type(s) of DNA recoveredss DNA 및 dsDNA

리소스

User-Developed Protocols (1)
The protocol has been used successfully for Cy3-, Cy5-, and biotin-labeling of cDNA from <50 ng of total RNA or poly A+ mRNA.
Kit Handbooks (1)
Safety Data Sheets (1)
Certificates of Analysis (1)

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
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
Can QIAquick Kits be used to clean up RNA samples?
Although it is possible to use the QIAquick Nucleotide Removal Kit and QIAquick PCR Purification Kit for RNA purification, the conditions are not optimized for RNA, nor are RNase-free conditions guaranteed. The RNeasy MinElute Cleanup Kit is recommended for clean-up, concentration and desalting of RNAs above 200 bases in length.
FAQ ID -490
Do CoralLoad dyes supplied in various QIAGEN PCR Kits interfere with downstream applications?

CoralLoad dyes supplied in PCR Kits such as, e.g., Taq, HotStarTaq, and TopTaq DNA Polymerase and TopTaq Master Mix do not interfere with most downstream enzymatic applications.

However, for reproducible results, purification of PCR products using the QIAquick or MinElute PCR Purification Kits prior to enzymatic manipulation is recommended.

 

 

FAQ ID -1745
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
Do you have a protocol for purification of DNA fragments from Cy3/Cy5 dye-label reactions?
Yes, please follow the Supplementary Protocol 'Purification of DNA fragments from dye-labeled reactions using the QIAquick PCR Purification Kit' (QQ06). 
FAQ ID -947
What is the composition of Buffer PB?
Buffer PB contains a high concentration of guanidine hydrochloride and isopropanol. The exact composition of Buffer PB is confidential. However, this buffer can be purchased separately: Buffer PB.
FAQ ID -2791
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 I have to remove the oil from my PCR reaction before using the QIAquick or MinElute PCR Purification Kit?
No - mineral oil will not affect the clean-up procedure with the QIAquick or MinElute PCR Purification Kit.
FAQ ID -575
Can I use the QIAquick PCR Purification Kit for restriction enzyme cleanup?

Yes. The QIAquick PCR Purification Kit has been used to clean up fragments between 100 bp and 10 kb from a wide range of enzymatic reactions, removing salts, buffers, enzymes, nucleotides, and primers smaller than 40 nucleotides. Reactions that can be cleaned up with the QIAquick PCR Purification Kit include restriction digests, random priming, ligase, kinase, phosphatase, nuclease, nick translation, and cDNA synthesis reactions.
For data and additional information, please see QIAGEN News article Issue No. 5, 1998 "Fast and efficient enzyme removal with QIAquick Spin kits."

FAQ ID -130
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