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Taq PCR Core Kit

用于常规和特殊的PCR，包含dNTP混合液

  • QIAGEN PCR缓冲体系最大限度地减少了PCR条件的优化
  • 独特的即用型PCR缓冲液，操作更快速简单
  • Q-Solution可帮助扩增GC含量高的模板
  • 提供多种规格的包装方便使用
Taq PCR Core Kit包含Taq DNA Polymerase、独特的QIAGEN PCR Buffer能最大限度地减少使用时对PCR条件的优化，以及dNTP混合液。同时还提供新型的辅助剂Q-Solution，有助于实现某些“困难”模板（比如，高GC含量的模板）的高效扩增。CoralLoad PCR Buffer（包括两种胶示踪染料）使PCR产物可以直接上样电泳。
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Product Cat. no. List price:
Taq PCR Core Kit (250 U)
250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2, dNTP Mix
 201223
1.465,00 kr.
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Taq PCR Core Kit (1000 U)
4 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2, dNTP Mix
 201225
4.955,00 kr.
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The Taq PCR Core Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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CoralLoad PCR Buffer。|适用于不同的镁离子浓度。|扩增困难样本。|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 PCR Core Kit在大量的PCR应用中有优秀的PCR性能，无需耗时的优化过程。该试剂盒包含Taq DNA Polymerase，一种高品质的重组酶适用于常规和特殊的PCR应用（参见"Tolerance of different primer Tm Values"和"Specific amplification of long PCR products"）。每批Taq DNA Polymerase受到全面地质量监控测试，包括严格的PCR特异性和重复性测试，在此测试中可从人类基因组DNA中扩增低拷贝靶（参见"Lot-to-lot reproducibility"）。试剂盒中还包含独特制备的QIAGEN PCR Buffer和CoralLoad PCR Buffer，能在最少的优化过程的PCR条件下，进行高特异性的PCR（参见"Wide annealing-temperature window"和"Tolerance to variable magnesium concentration"）。此外，CoralLoad PCR Buffer能使PCR产物直接上样到琼脂糖凝胶，操作简单，能更快得到结果。这次的PCR可通过使用试剂盒内的PCR辅助剂Q-Solution进行优化（参见"Amplification of difficult templates"）。

Taq DNA Polymerase规格

浓度：5单位/µl 
重组酶：有
底物类似物：dNTP、ddNTP、dUTP、biotin-11-dUTP、DIG-11-dUTP、荧光-dNTP/ddNTP 
延伸率：72°C下2–4 kb/分钟
半衰期：97°C下10分钟；94°C下60分钟
扩增效率：≥105倍
5'–>3'外切酶活性：有
Extra A辅助剂: 有
3'–>5'外切酶活性：无
污染核酸：无
污染RNA酶：无
污染蛋白酶：无
自吸泵活性：无

原理
Taq PCR Core Kit包含所有简单可靠的PCR所需的所有试剂，Taq DNA Polymerase、QIAGEN PCR Buffer、CoralLoad PCR Buffer、Q-Solution、dNTP Mix和MgCl2.
Taq DNA Polymerase

Taq DNA Polymerase是高品质的重组酶，适用于常规的和特殊的PCR应用（参见"Tolerance of different primer Tm values"和"Specific amplification of long PCR products"）。
QIAGEN PCR Buffer

研发新型的QIAGEN PCR Buffer通过减少使用时对PCR条件的优化，来节省时间和精力。QIAGEN PCR Buffer包括KCl和(NH4)2SO4。独特的缓冲体系有助于特异性PCR产物的扩增。在每个PCR循环的退火步骤中，该缓冲液具有特异性-非特异性引物的高比率结合。由于KCl和(NH4)2SO4独特的均衡结合，相比传统的PCR缓冲液，该PCR缓冲液提供大范围退火温度和Mg2+浓度条件，以及严格的引物退火条件。通过改变退火温度或Mg2+浓度，即可显著减少PCR优化过程，而且通常不需要优化（参见"Wide annealing temperature window"和"Tolerance to variable magnesium concentration"）。

CoralLoad PCR Buffer

CoralLoad PCR Buffer具有所有QIAGEN PCR Buffer的优点。此外，还可以直接将PCR反应物上样到琼脂糖凝胶上，无需额外单独的凝胶上样缓冲液。CoralLoad PCR Buffer与传统的QIAGEN PCR Buffer一样能提供高PCR特异性和最小的反应优化条件。而且该缓冲液还包含两种标记染料，橙色染料和红色染料，有助于消除DNA迁移距离并优化琼糖脂凝胶跑样时间（参见"CoralLoad PCR Buffer"）。该缓冲液优化了移液的可视性，并能够直接将PCR产物上样到凝胶上，便于操作。

Q-Solution

Q-Solution有利于通过修改DNA熔化行为扩增高GC含量的模板或具有高度二级结构的模板。使用该特殊的试剂通常能够进行或提高欠佳的PCR（参见"Amplification of difficult templates"）。不同于DMSO和其他的PCR辅助剂，Q-Solution用于各种引物模板的特定浓度，且无毒。

操作流程
Taq PCR Core Kit提供所有构建PCR反应体系和用于多重基于PCR应用的所有组件。该试剂盒提供经优化、易于使用、精简的实验方案，确保获得成功的PCR结果。该试剂盒中还包含一款独特的PCR辅助剂，Q-Solution，用于简化欠佳的PCR反应。
应用

Taq DNA Polymerase用于常规的和特殊的应用，包括：

  • 常规PCR
  • RT-PCR
  • 筛选 
  • 基于PCR的DNA指纹图谱分析（VNTR、STR和RAPD）
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Feature
Specifications
Applications PCR, RT-PCR, DNA fingerprinting
dNTP's included Yes
Enzyme activity 5' -> 3' exonuclease activity
Mastermix No
Reaction type PCR amplification
Real-time or endpoint Endpoint
Sample/target type Genomic DNA and cDNA
Single or multiplex Single
With/without hotstart Without hotstart

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Kit Handbooks
1
Taq PCR Handbook - (EN)
For standard and specialized PCR applications with minimal optimization
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Safety Data Sheets
1
Safety Data Sheets
Download Safety Data Sheets for QIAGEN product components.
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Images
Ready-to-load PCR buffer
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.
A Wide Annealing-Temperature Window
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 with Q-Solution
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.
Tolerance of different primer Tm values.
The human single-copy cystic fibrosis gene was amplified usingTaq 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.
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.
Specific amplification of long PCR products.
Three different-sized products from human genomic DNA were amplified using either Taq DNA Polymerase and the 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.
NH4+ and K+ cations in QIAGEN PCR buffers increase specific primer annealing
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 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.