Type-it Mutation Detect PCR Kit

用于对突变进行准确、可靠的多重PCR分析

  • 有效、可重复的多重突变分析
  • 无需优化即可建立多重PCR反应
  • 高特异性、灵敏的同时扩增所有片段
  • 优化的操作流程,得到快速、可靠的结果
  • 简单的操作指南,用于多种下游分析平台

Type-it Mutation Detect PCR Kit适用于快速、可靠的检测缺失、插入和异位等突变。该试剂盒基于高度特异的HotStarTaq Plus DNA Polymerase和获得专利的缓冲液系统,从而确保对突变位点进行可靠多重PCR扩增,无需优化。后续分析简单方便,可在琼脂糖凝胶、自动化电泳仪以及高分辨率毛细管测序仪上进行。

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Type-it Mutation Detect PCR Kit (200)
For 200 x 25 μl reactions: Type-it Multiplex PCR Master Mix, 5x Q-Solution, RNAse-Free Water, and 10x CoralLoad Dye
206343
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The Type-it Mutation Detect 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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CoralLoad Dye.|Sensitive detection of a mutated cancer-related gene.|Superior preamplification of SNPs.|Successful multiplex PCR-based genotyping analysis.|Stable and efficient annealing.|
[A] CoralLoad Dye contains gel-tracking dyes, enabling immediate gel loading of PCR and [B] easy visualization of DNA migration.|The indicated amounts of DNA extracted from a lymphoma-related cell line (Ramos) were spiked into human leukocyte DNA, as indicated in the figure, and the mutated Ramos target was detected along with 2 internal controls. Using the Type-it Mutation Detect Kit, the mutated 357 bp gene was detected even when only 25 pg of DNA (4 cells) was present. The lymphoma-relevant mutation could not be detected when using preoptimized conditions (Mg2+ and polymerase) with a hot-start DNA polymerase from Supplier I — even when using 25 ng of DNA (4000 cells). Electrophoresis was performed on a 1.3% agarose gel. M: 100 bp ladder.|Two different PCR systems (12-plex and 5-plex), both including the SNP of interest, were amplified using the standard protocol for the Type-it Mutation Detect PCR Kit (Q) or using preoptimized conditions with a hot-start DNA polymerase from Supplier A. With Type-it Mutation Detect Kits, all fragments of interest were amplified without any optimization of PCR parameters and could successfully be used as input for the SNaPshot PCR system. Even after optimization of Mg2+ concentration to 4 mM and varying the polymerase concentrations, amplification using the kit from Supplier A resulted in missing fragments. Electrophoresis was performed on the QIAxcel System. The first and the last bands on each lane are Upper Alignment Marker and Lower Alignment Marker, respectively. M: Alignment marker.|The Type-it Microsatellite PCR Kit is based on proprietary QIAGEN multiplex technology and provides a simple procedure for reliable genotyping results. In contrast to current methods, the need for optimization of PCR parameters such as Mg2+ concentration and cycling conditions is virtually eliminated.|Unique Type-it Mutation Detect PCR Buffer promotes stable and efficient 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 Plus DNA Polymerase.|
Performance
Type-it Mutation Detect PCR Kit优于其他供应商的试剂盒,确保可靠的突变分析。该试剂盒专门研发并经功能验证,可用于对缺失或插入等突变进行多重PCR分析,也可检测遗传修饰的有机体或微生物(参见"Sensitive detection of a mutated cancer-related gene)。该试剂盒也适合用作商业化SNP检测PCR体系的预扩增方法,如Applied Biosystems提供的SNaPshot体系(参见"Superior preamplification of SNPs")。
Principle

Type-it Mutation Detect PCR Kit基于专有的QIAGEN Multiplex Technology,以即用型预混液形式提供。预混液含有浓度经优化的HotStarTaq Plus DNA Polymerase、MgCl2、dNTPs和一种创新的PCR缓冲液,专用于对突变进行多重PCR检测,或是对基因组SNP位点进行预扩增。该试剂盒还包含创新的添加剂Factor MP,以及平衡比例的盐和添加剂。这些组分使反应中的所有引物都具有相当的退火和延伸效率(参见"Stable and efficient annealing"),可平行地对所有片段进行可靠的高产量多重扩增。该试剂盒包括专门用于检测突变的操作流程,以及对模板量、循环数、PCR条件和不同下游分析平台仪器细节的建议,分析平台包括琼脂糖凝胶、毛细管测序仪、Agilent Bioanalyze和QIAxcel Advanced System。

预混液中所有组分配合专有的操作流程,可平行地对所有片段进行高特异性扩增。后续分析简单、直接,可在琼脂糖凝胶、自动化电泳仪上进行,也可进行高分辨率毛细管测序。

Type-it Mutation Detect PCR Buffer

Type-it Mutation Detect PCR Buffer确保高级别多重实验中的所有扩增子具有相当的扩增效率,可整合多个突变靶点,不会丢失任何突变靶点的扩增效率。与常规PCR试剂不同,Type-it Mutation Detect PCR Buffer含有特别研发的、比例平衡的盐和添加剂,确保反应中所有引物的退火和延伸具有同等的效率。KCl和(NH4)2SO4独特的结合比例,使PCR缓冲液与常规PCR缓冲液相比,可在更宽范围的退火温度和Mg2+浓度下提供严格的引物退火条件。极大的减少通过改变退火温度或Mg2+浓度的PCR优化过程,有时甚至不需要。通常使用的多重PCR优化步骤几乎不需要。缓冲液中还含有合成的Factor MP(参见"Stable and efficient annealing"),不论引物序列如何都可进行有效的引物退火和延伸。Factor MP可提高DNA模板处的引物浓度,稳定特定结合的引物。

Q-Solution

Type-it Mutation Detect PCR Kit提供Q-Solution。这种创新的PCR添加剂通过修饰DNA的熔解行为,便于扩增难扩增的模板。使用这种独特的试剂常常能完成或改进不理想的PCR。与DMSO和其他PCR添加剂不同,Q-Solution可在多种引物-模板体系中使用特定工作浓度,而不会产生毒性作用。

CoralLoad Dye

Type-it Mutation Detect PCR Kit提供CoralLoad Dye(参见"CoralLoad Dye"),这种染料含有一种胶上样试剂和两种胶示踪染料,提高移液的可见性,便于估计DNA迁移距离及优化琼脂糖凝胶电泳时间。在多重PCR反应中使用CoralLoad Dye,扩增产物可直接上样到琼脂糖凝胶或QIAxcel Advanced System中,无需提前加入上样缓冲液。CoralLoad染料不会干扰大部分下游酶应用。但是,为获得可重复性的结果,建议在进行酶处理前纯化PCR产物。                                                    如果使用毛细管测序仪进行检测,不能使用CoralLoad Dyes。

Procedure

Type-it Mutation Detect PCR Kit包括专有的、特异性的操作流程,可同时扩增和后续检测含突变的位点或SNPs,保证在常规分析中获得可靠结果,也可用于新检测的建立。可在室温下建立反应,确保更大的便利性和易用性。

快速、简单的方法,获得可靠的基因分型结果

Type-it Mutation Detect PCR Kit确保快速、简单的多重分析构建,以获得准确、可重复的基因分型结果。无论检测易位、缺失、插入或预扩增基因组位点用于SNP分析,如SNaPshot(Applied Biosystems),只需加入模板和引物后,根据优化方案开始热循环程序即可。反应混合物含有突变特异性多重PCR所需的所有试剂,与目前的其他方法相比,无需冗长的优化步骤即可获得准确的结果(参见"Successful multiplex PCR-based genotypic analysis")。

简单的操作流程,节约时间和成本

有些研究需要分析疾病相关的某个基因的很多不同突变(如缺失、易位或插入),如果进行单重或低级别PCR分析,需要众多的PCR反应,增加时间和成本。Type-it Mutation Detect PCR Buffer确保高级别多重实验中的所有扩增子具有相当的扩增效率,可整合多个突变靶点,大大节约时间和成本(参见"Sensitive detection of a mutated cancer-related gene")。此试剂盒含有专门的、应用特异性的操作流程,可同时扩增和后续检测含突变的位点或SNPs,保证在常规分析中获得可靠结果,也可用于新检测的建立。

可靠的SNP预扩增 

Type-it Mutation Detect PCR Kit非常适合对SNPs进行多重PCR预扩增(如来自Applied Biosystems的SNaPshot系统)。此试剂盒优于来自其他供应商的试剂盒,可持续获得可靠的结果,无需繁琐的优化步骤(参见"Superior preamplification of SNPs")。

Applications

Type-it Mutation Detect PCR Kit可用于分析缺失、插入、重复和易位的突变,以及进行SNP预扩增(如SNaPshot Multiplex Kit)。该试剂盒可用于多种研究领域:

  • 疾病基因位点的分型
  • 转基因生物分析
  • 转基因植物或动物的分型
Feature
Specifications
Applications Multiplex PCR, detection of mutations, preamplification of SNPs
Mastermix Yes
Product use Functionally validated and developed for reliable mutation analyis
Reaction type PCR amplification
Real-time or endpoint Endpoint
Sample/target type Genomic DNA
Single or multiplex Multiplex
With/without hotstart With

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Brochures & Guides
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Kit Handbooks
1
For optimization-free multiplex PCR-based mutation detection or preamplification of SNPs
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