For standard and specialized PCR applications
- QIAGEN PCR Buffer for minimal optimization
- Additional ready-to-load PCR buffer for faster handling
- Q-Solution for amplification of GC-rich templates
- Choice of formats for convenience and ease of handling
Taq DNA Polymerase is supplied with the unique QIAGEN PCR Buffer that minimizes the need for optimization of PCR parameters, as well as Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC rich) templates. In addition, CoralLoad PCR Buffer (containing two gel-tracking dyes) is also provided, enabling immediate loading of PCR products.
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Taq DNA Polymerase (250 U)
250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
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201203
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Taq DNA Polymerase (1000 U)
4 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
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201205
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Taq DNA Polymerase (5000 U)
20 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
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201207
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Taq DNA Polymerase Kit (25000 U)
100 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
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201209
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The Taq DNA Polymerase is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
CoralLoad PCR Buffer。|可用于不同镁离子浓度。|扩增困难模板。|对不同的引物Tm值的适应度。|孔间可重复性。|特异性扩增获得长片段PCR产物。|退火时引物结合的特异性增强。|
[A] CoralLoad染料包含两种凝胶示踪染料,能够便利的进行凝胶上样,[B] 以及清楚的看到DNA的迁移。|使用QIAGEN PCR Buffer和Taq DNA Polymerase (QIAGEN),在图示的Mg2+浓度下进行PCR扩增。使用另一家供应商 (Supplier AII) 的PCR缓冲液和Taq聚合酶同时进行相同的PCR。使用QIAGEN的缓冲液和酶可在各种情况下完成单拷贝人朊病毒蛋白基因的扩增。M:分子量标准。|使用QIAGEN PCR Buffer和Taq DNA Polymerase,在1x Q-Solution存在 (+) 或不存在 (–) 的情况下,对两种不同的引物-模板系统进行扩增,重复两次。Q-Solution可实现复杂模板的特异性扩增。[A] 人血管紧张素受体II基因;[B] 小鼠蛋白激酶C基因;M:分子量标准。|采用图示的退火温度,使用Taq DNA Polymerase和QIAGEN PCR Buffer扩增单拷贝的人囊性纤维化基因。采用的引物为22mer,Tm为57.5°C (GC含量:54.5%),以及32mer,Tm为85.2°C (GC含量:78%)。取100 µl反应产物的10%上样分析。M:分子量标准。|从30 ng、3 ng和300 pg人基因组DNA中扩增单拷贝的囊性纤维化基因片段,这些基因组DNA分别对应104、103和102拷贝的目标模板。使用3个不同批次的Taq DNA Polymerase进行扩增,取等体积的PCR产物在1%的琼脂糖凝胶上进行分析。M:分子量标准。|使用Taq DNA Polymerase和QIAGEN PCR Buffer (QIAGEN) 或者另一家供应商的Taq聚合酶和缓冲液 (Supplier AII) 扩增3种不同大小的人基因组DNA产物。取各反应产物的10%上样于凝胶上进行分析。两次PCR重复扩增的结果如图所示。M:分子量标准。|QIAGEN PCR缓冲液中的铵态氮和钾离子能够在退火时促进引物的特异性结合。K+结合到DNA主链上的磷酸基团(P–)上,稳定结合到模板上的引物。在热循环中,NH4+以铵离子和氨的形式存在,可以与碱基(B)之间的氢键发生相互作用,使错配碱基间的不稳定的氢键容易断开。两种阳离子的共同作用,在广泛的温度范围内保持特异性结合比例高。|
性能
Taq DNA Polymerase outperformed kits tested from other suppliers and delivers robust PCR performance in a wide range of PCR conditions, without the need for time-consuming optimization (see figures "Tolerance of different primer Tm Values" and "Specific amplification of long PCR products"). Every lot of Taq DNA Polymerase is subjected to a comprehensive range of quality control tests, including a stringent PCR specificity and reproducibility assay in which low-copy targets are amplified from human genomic DNA (see figure "Lot-to-lot reproducibility"). The unique formulation of QIAGEN PCR Buffer and CoralLoad PCR Buffer, also provided with the kit, enable highly specific PCR in a variety of PCR conditions with minimal optimization requirements (see figures "Wide annealing-temperature window" and "Tolerance to variable magnesium concentration"). In addition, CoralLoad PCR Buffer enables immediate loading of PCR products onto an agarose gel for even easier handling and faster results. Suboptimal PCR can be improved using Q-Solution, a PCR additive, also provided with the kit (see figure "Amplification of difficult templates").
Taq DNA Polymerase specifications
Concentration: 5 units/µl Recombinant enzyme: Yes Substrate analogs: dNTP, ddNTP, dUTP, biotin-11-dUTP, DIG-11-dUTP, fluorescent-dNTP/ddNTP Extension rate: 2–4 kb/min at 72°C Half-life: 10 min at 97°C; 60 min at 94°C Amplification efficiency: ≥105 fold 5'–>3' exonuclease activity: Yes Extra A addition: Yes 3'–>5' exonuclease activity: No Contaminating nucleases: No Contaminating RNases: No Contaminating proteases: No Self-priming activity: No
原理
Taq DNA Polymerase is a high-quality recombinant enzyme that is suitable for general and specialized PCR applications (see figures "Tolerance of different primer Tm Values" and "Specific amplification of long PCR products").
QIAGEN PCR Buffer
Innovative QIAGEN PCR Buffer has been developed to save time and effort by reducing the need for PCR optimization. QIAGEN PCR Buffer contains both KCl and (NH4)2SO4(see figure "Increased specificity of primer annealing"). This unique buffer facilitates the amplification of specific PCR products. During the annealing step of every PCR cycle, the buffer allows a high ratio of specific-to-nonspecific primer binding. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced and often not required (see figures "Wide annealing temperature window" and "Tolerance to variable magnesium concentration").
CoralLoad PCR Buffer
CoralLoad PCR Buffer has all the advantages of QIAGEN PCR Buffer. In addition, it can also be used to directly load the PCR reaction onto an agarose gel — separate addition of a gel loading buffer is not required. CoralLoad PCR Buffer provides the same high PCR specificity and minimal reaction optimization as the conventional QIAGEN PCR Buffer. Additionally, it contains two marker dyes — an orange dye and a red dye — that facilitate estimation of DNA migration distance and optimization of agarose gel run time (see figure "CoralLoad PCR Buffer"). The buffer ensures improved pipetting visibility and enables direct loading of PCR products onto a gel, for enhanced convenience.
Q-Solution
Q-Solution facilitates amplification of GC-rich templates or templates with a high degree of secondary structure by modifying the melting behavior of DNA. Use of this unique reagent often enables or improves suboptimal PCR (see figure "Amplification of difficult templates"). Unlike DMSO and other PCR additives, Q-Solution is used at a defined working concentration with any primer–template system and is not toxic.
操作流程
Taq DNA Polymerase ensures highly specific PCR for a range of applications — with minimal optimization of PCR parameters. The streamlined, easy-to-follow protocol provided with the kit simplifies PCR setup. For added convenience and easier handling, CoralLoad PCR Buffer is provided. PCR products can be directly loaded onto a gel without the addition of a loading dye. To ensure success with GC-rich templates, the PCR enhancer Q-Solution is included.
应用
Taq DNA Polymerase is used for standard and specialized applications, including:
- General PCR
- RT-PCR
- Screening
- PCR-based DNA fingerprinting (VNTR, STR, and RAPD)
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For standard and specialized PCR applications with minimal optimization
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CoralLoad PCR Buffer.
[A] CoralLoad Concentrate contains two gel-tracking dyes, enabling immediate gel loading of PCR products for [B ] easy visualization of DNA migration.
Tolerance to variable magnesium concentration.
PCR amplification at the indicated Mg2+ concentrations using QIAGEN PCR Buffer and Taq DNA Polymerase (QIAGEN). The same PCR was performed in parallel using a PCR buffer and Taq polymerase from another supplier (Supplier AII). The single-copy human prion protein gene was amplified successfully in each case using the buffer and enzyme from QIAGEN. M: markers.
Amplification of difficult templates.
Two different primer-template systems were amplified in duplicate using QIAGEN PCR Buffer and Taq DNA Polymerase in the absence (–) or presence (+) of 1x Q-Solution. Q-Solution enables specific amplification of difficult templates. [A] human angiotensin receptor II gene; [B] mouse protein kinase C gene; M: markers.
Tolerance of different primer Tm values.
The human single-copy cystic fibrosis gene was amplified with Taq DNA Polymerase and QIAGEN PCR Buffer using the indicated annealing temperatures. Primers employed were a 22mer with a Tm of 57.5°C (GC content: 54.5%) and a 32mer with a Tm of 85.2°C (GC content: 78%). For analysis, 10% of a 100 µl reaction was loaded. M: markers.
Lot-to-lot reproducibility.
A fragment of the single-copy gene for cystic fibrosis was amplified from 30 ng, 3 ng, and 300 pg human genomic DNA corresponding to 104, 103, and 102 copies of target template, respectively. Three different lots of Taq DNA Polymerase were used and equal volumes of the PCR product were analyzed on a 1% agarose gel. M: markers.
Specific amplification of long PCR products.
Three different-sized products from human genomic DNA were amplified using either Taq DNA Polymerase and QIAGEN PCR Buffer (QIAGEN), or Taq polymerase and a buffer from another supplier (Supplier AII). For analysis, 10% of each reaction was loaded on the gel. Results from duplicate PCR amplifications are shown. M: markers.
Increased specificity of primer annealing.
Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K+ binds to the phosphate groups (P–) on the DNA backbone, stabilizing the annealing of the primers to the template. NH4+, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing the weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains the high ratio of specific-to-nonspecific primer-template binding over a wide temperature range.
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