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)
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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
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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:
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
5'–>3'外切酶活性:有
Extra A辅助剂: 有
3'–>5'外切酶活性:
污染核酸:
污染RNA酶:
污染蛋白酶:
自吸泵活性:

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+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
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
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Kit Handbooks
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For fast and efficient multiplex PCR without optimization
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