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QIAGEN Multiplex PCR Kit

用于高特异性和灵敏度的多重PCR，无需优化

  • 无需优化步骤
  • 高特异性和高灵敏度，热启动
  • 高度适合用于各种类型的多重PCR应用
  • 使用简单，经济实惠

QIAGEN Multiplex PCR Kit提供方便的即用型预混液。QIAGEN Multiplex PCR Master Mix包含HotStarTaq DNA Polymerase和含有新型合成因子MP的独特PCR缓冲液。配合优化的盐浓度，MP因子可以促进引物与模板的特异性结合，使反应体系中所有的引物都能有效延伸，无需额外优化。该试剂盒还包含新型的辅助剂Q-Solution，有助于实现“困难”模板（比如，GC含量高的模板）的高效扩增。

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QIAGEN Multiplex PCR Kit (100)
For 100 x 50 µl multiplex PCR reactions: 2x QIAGEN Multiplex PCR Master Mix (providing a final concentration of 3 mM MgCl2, 3 x 0.85 ml), 5x Q-Solution (1 x 2.0 ml), RNase-Free Water (2 x 1.7 ml)
 206143
€ 264,00
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QIAGEN Multiplex PCR Kit (1000)
For 1000 x 50 µl multiplex PCR reactions: 2x QIAGEN Multiplex PCR Master Mix (providing a final concentration of 3 mM MgCl2, 1 x 25 ml), 5x Q-Solution (1 x 10 ml), RNase-Free Water (1 x 20 ml)
 206145
€ 2.307,00
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The QIAGEN Multiplex PCR 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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Successful 16-plex PCR.|Genotyping transgenic mice.|Successful microsatellite analysis.|Stable and efficient primer annealing.|
Multiplex PCR of 16 targets (99–955 bp) was carried out for 35 cycles using standard conditions for the QIAGEN Multiplex PCR Kit, without further optimization or using a variety of conditions with a hot-start DNA polymerase from Supplier AII. [A] Comparison using 2.5 units per 50 µl reaction of the hot-start DNA polymerase from Supplier AII and with the indicated Mg2+ concentrations. [B] Comparison using the optimized Mg2+ concentration (3.5 mM) for the hot-start DNA polymerase from Supplier AII (part A) and the indicated amounts of enzyme per 50 µl reaction. Successful results were ensured with the QIAGEN Multiplex PCR Kit. M: markers.|Transgenic mice were screened using the QIAGEN Multiplex PCR Kit and sets of 3 primers to distinguish wild-type (wt), heterozygous mutant (ht), and homozygous mutant (hm) mice. [A] Using a primer set for the recombination activating gene 2 locus. [B] Using a primer set for the interferon-γ gene locus. M: markers. (Data kindly provided by S. zur Lage and S. Weiss, National Research Center for Biotechnology, Braunschweig, Germany.)|Analysis of microsatellite loci D3S1358, TH01, D21S11, D18S51, and Penta E was carried out using 1 ng of K562 human genomic DNA and fluorescein-labeled primers. Reactions were analyzed on the ABI PRISM 377 Sequencer. Top: The QIAGEN Multiplex PCR Kit ensured high sensitivity and uniform signal intensity. Bottom: Results using a hot-start DNA polymerase from Supplier AII.|The Multiplex PCR Buffer ensures stable and efficeint primer annealing. [A] NH4+ ions prevent nonspecific primers from annealing to the template. [B] Synthetic factor MP, an innovative PCR additive, increases the local concentration of primers at the template. Together with K+ and other cations, synthetic factor MP stabilizes specifically bound primers, allowing efficient primer extension by HotStarTaq DNA Polymerase.|
Performance

QIAGEN Multiplex PCR Kit可确保高特异性和灵敏度的多重PCR扩增（参见"Successful 16-plex PCR "）。该试剂盒能成功应用于各种多重应用，如转基因生物分型（参见"Genotyping transgenic mice"）和微卫星分析（参见"Successful microsatellite analysis "）。该混合物包含HotStarTaq DNA Polymerase，用于高效的平行扩增多个靶。扩增效率经新型的PCR缓冲液进一步优化，该缓冲液也包含在混合物中。独特的缓冲液确保了在广泛的PCR条件下PCR的特异性，无需优化步骤。试剂盒还包含辅助剂Q-Solution，用于扩增高GC含量的模板，优化PCR反应。

HotStarTaq 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

Principle

QIAGEN Multiplex PCR Kit是首个专为多重PCR研发的试剂盒，有即用型混合液规格。QIAGEN Multiplex PCR Master Mix包含预优化浓度的HotStarTaq DNA Polymerase和MgCl2、dNTPs和新型的PCR缓冲液专为多重PCR研发。该试剂盒在首次尝试多重PCR即获得成功。由于试剂盒内独特的预优化的试剂无需优化反应条件（比如，引物浓度，Mg2+和Taq DNA聚合酶）和循环参数。

HotStarTaq DNA Polymerase

HotStarTaq DNA Polymerase对 Taq DNA聚合酶进行改善，在室温下无聚合酶活性。这可以防止非特异性退火引物和在低温条件下的PCR设置和初始的PCR循环中形成引物二聚体。在95°C下，经15分钟诱导可激活HotStarTaq DNA Polymerase，可纳入各种现有的热循环程序中。

多重PCR缓冲液

这特殊的缓冲液包含优化的由K+和NH4+以及独特的PCR辅助剂、MP因子可增加模板引物的局部浓度。配合K+和其他阳离子、MP因子稳定特异性结合的引物，并能够用HotStarTaq DNA Polymerase进行高效的引物延伸（参见"Stable and efficient primer annealing"）。通过在每个PCR循环中高比率的特异性-非特异性引物结合，该新型的缓冲液维持每个PCR循环的特异性扩增。由于KCl和(NH4)2SO4独特的平衡结合，相比传统的PCR缓冲液，该缓冲液能在更广泛的退火温度和Mg2+浓度下，提供严格的引物退火条件。通过改变退火温度或Mg2+浓度可优化PCR，因此通常将优化过程最小化或无需要优化。

Q-Solution

Q-Solution是一种新型的PCR辅助剂，有助于扩增由DNA融化特性修改的困难模板，同时配合HotStarTaq DNA Polymerase提供。该独特的试剂可优化高度二级结构或富含GC的模板所致的效果欠佳的PCR。不同于其他常用的PCR辅助剂如DMSO，Q-Solution只用于一种工作浓度，并保证无毒和PCR纯度好。

Procedure

QIAGEN Multiplex PCR Kit含有即用型、预优化的混合液，非常方便。使用混合液节省时间、简化反应体系构建流程，并通过消除移液失误和污染，增加可靠性，减少移液步骤和繁琐的计算。只需加入引物和模板来制备最终扩增混合液。混合液可在2–8°C储存，可快速设置多重PCR检测。该试剂盒提供的精简的实验方案确保快速简单的PCR设置。可在室温下设置反应，简单方便，易于使用。HotStarTaq DNA Polymerase可在95°C下孵育15分钟激活，因此可整合入各种现有的热循环流程中。

Applications

QIAGEN Multiplex PCR Kit高度适用于各种多重应用，包括：

  • 转基因生物的分型和分析
  • 微卫星的扩增和分析
  • 细菌和病毒的分型和检测
  • 用于SNP分析的多个DNA扩增
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Feature
Specifications
Applications PCR, RT-PCR, multiplex PCR, typing, detection
Enzyme activity 5' -> 3' exonuclease activity
Mastermix Yes
Reaction type PCR amplification
Real-time or endpoint Endpoint
Sample/target type Genomic DNA and cDNA
Single or multiplex Multiplex
With/without hotstart With hotstart

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Brochures & Guides
2
Analyzing Genetic Differences - (EN)
Second edition — innovative tools
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(EN) - Maximizing PCR and RT-PCR success — Third Edition
Addressing critical factors and new solutions
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Kit Handbooks
1
(EN) - QIAGEN Multiplex PCR Handbook
For fast and efficient multiplex PCR without optimization
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Safety Data Sheets
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Safety Data Sheets
Download Safety Data Sheets for QIAGEN product components.
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Images
Successful 16plex PCR Using the QIAGEN Multiplex PCR Kit Multiplex
Successful 16-plex PCR.
Multiplex PCR of 16 targets (99–955 bp) was carried out for 35 cycles using standard conditions for the QIAGEN Multiplex PCR Kit, without further optimization or using a variety of conditions with a hot-start DNA polymerase from Supplier AII. [A] Comparison using 2.5 units per 50 µl reaction of the hot-start DNA polymerase from Supplier AII and with the indicated Mg2+ concentrations. [B] Comparison using the optimized Mg2+ concentration (3.5 mM) for the hot-start DNA polymerase from Supplier AII (part A) and the indicated amounts of enzyme per 50 µl reaction. Successful results were ensured with the QIAGEN Multiplex PCR Kit. M: markers.
Genotyping Transgenic Mice
Genotyping transgenic mice.
Transgenic mice were screened using the QIAGEN Multiplex PCR Kit and sets of 3 primers to distinguish wild-type (wt), heterozygous mutant (ht), and homozygous mutant (hm) mice. [A] Using a primer set for the recombination activating gene 2 locus. [B] Using a primer set for the interferon-γ gene locus. M: markers. (Data kindly provided by S. zur Lage and S. Weiss, National Research Center for Biotechnology, Braunschweig, Germany.)
Microsatellite Analysis with Optimized QIAGEN Protocol
Successful microsatellite analysis.
Analysis of microsatellite loci D3S1358, TH01, D21S11, D18S51, and Penta E was carried out using 1 ng of K562 human genomic DNA and fluorescein-labeled primers. Reactions were analyzed on the ABI PRISM 377 Sequencer. Top: The QIAGEN Multiplex PCR Kit ensured high sensitivity and uniform signal intensity. Bottom: Results using a hot-start DNA polymerase from Supplier AII.
Unique Type-it Microsatellite PCR Buffer Promotes Stable and Efficient Annealing
Stable and efficient primer annealing.
The Multiplex PCR Buffer ensures stable and efficeint primer annealing. [A] NH4+ ions prevent nonspecific primers from annealing to the template. [B] Synthetic factor MP, an innovative PCR additive, increases the local concentration of primers at the template. Together with K+ and other cations, synthetic factor MP stabilizes specifically bound primers, allowing efficient primer extension by HotStarTaq DNA Polymerase.