Type-it Fast SNP Probe PCR Kit

For accurate and reliable SNP genotyping using TaqMan® or TaqMan® MGB probes

  • Validated using TaqMan® SNP Genotyping Assays
  • Automated allele calling and tight fluorescence clusters
  • Suitable for difficult SNP loci or very low template amounts
  • Up to 40% time savings due to fast cycling procedure
The Type-it Fast SNP Probe PCR Kit is based on highly specific HotStarTaq Plus DNA Polymerase and a newly developed buffer system, both of which enable reliable and clear allelic discrimination. The combination of all components provided in the master mix result in improved accuracy and highly specific probe binding. The Type-it Fast SNP Probe PCR Kit is validated using TaqMan® SNP Genotyping Assays and enables reproducible SNP genotyping — even with difficult SNP loci (e.g., GC rich) or low amounts of starting template.
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产品 货号 目录价:
Type-it Fast SNP Probe PCR Kit (800)
For 800 x 25 μl reactions: 6 x 1.7 ml of 2x Type-it Fast SNP Probe PCR Master Mix, 5x Q-Solution, RNase-Free Water
206045
询价
Type-it Fast SNP Probe PCR Kit (4000)
For 4000 x 25 μl reactions: 2 x 25 ml of 2x Type-it Fast SNP Probe PCR Master Mix, 5x Q-Solution, RNase-Free Water
206047
询价

Type-it Fast SNP Probe PCR Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


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成功的自动化等位基因信号分析。|高度特异性的探针结合。|基因分型工作流程。|使用少量模板进行可靠的SNP基因分型。|退火过程中快速循环的机制。|
使用6个不同的TaqMan MGB-based SNP基因分型实验和来自60个样本的1 ng基因组DNA对一组不同的DNA进行平均自动化等位基因信号率分析。在384孔板中以5 μl反应体系对指定产物进行PCR。在Applied Biosystems 7900HT仪器上进行等位基因区分读板。Type-it Fast SNP Probe PCR Kit可持续获得最高的信号率和最低的错误率。|由于错配探针的非特异性结合,来自其他供应商的用于SNP基因分型的缓冲液经常不能很好分离等位基因簇。Type-it Fast SNP Probe Buffer System通过缩小熔化温度范围,可提高分辨率,减少错配探针的非特异性结合。可对等位基因簇进行广泛的和明确的分离。|Type-it Fast SNP Probe PCR Master Mix含有高度特异性的HotStarTaq Plus Polymerase,可在室温下快速、简单地构建反应体系,即使模板量很少也能确保高度特异的扩增。|使用[A] Type-it Fast SNP Probe PCR Kit或[B]来自供应商A的基因分型预混液对80个不同基因组DNA(每5 μl反应有10 ng或1 ng)进行等位基因区分。即使使用少量模板,Type-it Fast SNP Probe PCR Master Mix的表现都优于来自供应商A的预混液,获得紧密的等位基因和可靠的结果。使用TaqMan SNP基因分型实验对rs 951134(aryl-hydrocarbon receptor repressor gene)和2个无模板对照(NTCs)进行PCR,在Applied Biosystems 7900HT仪器上进行分析。黑色:NTCs;蓝色:T等位基因纯合子(FAM荧光);绿色:杂合子样本;红色:A等位基因纯合子(VIC荧光)。数据以98%信度进行分析。|[A]专门的PCR缓冲液含有Q-Bond,可以加速退火步骤,将时间减少到5秒。Q-Bond提高Taq DNA聚合酶对短的单链DNA片段的亲和力,减少了引物退火所需时间。[B]没有Q-Bond时,引物和聚合酶依次结合到模板上,增加了退火步骤的持续时间。|
性能

The Type-it Fast SNP Probe PCR Kit consistently ensures highly accurate SNP genotyping. 

Highly stringent and specific binding of the allele-specific probe 

The Type-it Fast SNP Genotyping PCR Master Mix is based on highly specific HotStarTaq Plus DNA Polymerase and a newly developed SNP genotyping PCR buffer system, both of which enable highly specific probe binding and consistently strong fluorescent signals. Compared to other commercially available SNP genotyping master mix chemistries, wider and clearer separation of allele clusters is obtained (see figure "Highly specific probe binding").

High call rates even for low template amounts

Even with challenging targets or difficult SNP loci, the Type-it Fast SNP Probe PCR Kit provides reliable SNP genotyping — well-separated allele clusters and tight clustering of identical alleles are achieved, resulting in high call rates  (see figure "Successful automated allele calling"). The Type-it Fast SNP Probe PCR Kit outperformed kits tested from other suppliers, ensuring tight clustering of alleles — even with 1 ng of template DNA (see figure "Reliable SNP genotyping with small amounts of template").

原理

The Type-it Fast SNP Probe PCR Kit is available in a convenient master mix format consisting of highly specific HotStarTaq Plus Polymerase and a unique SNP genotyping buffer system designed for fast and efficient amplification. The unique kit components provide tight allele clustering and outstanding separation to ensure high call rates, enabling reproducible and accurate results (see table).

High specificity and sensitivity

HotStarTaq Plus DNA Polymerase, provided in the master mix, which ensures highly specific amplification, even with low template amounts. HotStarTaq Plus DNA Polymerase is provided in an inactive state with no polymerase activity at ambient temperatures. This prevents the formation of misprimed products and primer dimers at low temperatures. 

Unique buffer system

The Type-it Fast SNP PCR Buffer, also included in the master mix, is specifically designed for fast-cycling SNP genotyping using sequence specific 5'-nuclease probes. The unique composition of the Type-it Fast SNP PCR Buffer ensures highly stringent and specific binding of the allele-specific probe (match probe). This is due to the altered melting behaviour of the probes resulting in a narrower probe melting temperature window. Based on the original QIAGEN PCR Buffer, this innovative formulation enables a high ratio of specific-to-nonspecific primer binding during the short annealing step of every PCR cycle. 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.

Additives in the Type-it Fast SNP PCR buffer, such as Q-Solution, provide reaction conditions for amplification of difficult genomic regions and difficult SNP loci. Innovative Q-Bond technology (for fast cycling) allows SNP genotyping results to be achieved faster (see figure "Mechanism of fast cycling during annealing"). The master mix also contains ROX dye at a concentration compatible with SNP genotyping on all instruments from Applied Biosystems. However, ROX dye is also compatible with other instruments not requiring ROX as a passive reference dye (see table). 

Kit features
Kit contents Features
2x Master Mix format* Developed for SNP genotyping with TaqMan® MGB probes
Suitable for use with all cyclers suitable for SNP genotyping
HotStarTaq Plus DNA Polymerase Fast and easy reaction setup at room temperature
Highly specific amplification, even with low template amounts
Type-it Fast SNP Probe PCR Buffer Wide separation of genotype clusters
Tight allele clusters and high allele calling rates
Increased specificity in probe binding
Fast cycling due to Q-Bond Molecule
Q-Solution Tighter clusters and higher signals in scatter plot analysis for highly GC-rich SNP loci
Can further improve suboptimal allele calling
* Includes ROX reference dye, dNTPs, and MgCl2.
操作流程

The Type-it Fast SNP Probe PCR Kit is functionally verified with commercially available SNP genotyping assays and is compatible with TaqMan® MGB Probes, as well as user-developed probe-based assays consisting of TaqMan® MGB, TaqMan®, or other dual-labeled probes. The kit includes streamlined, preoptimized protocols for fast and reliable analysis.

Fast-cycling procedure for straightforward results

The Type-it Fast SNP Probe PCR Master Mix provides reaction conditions for fast-cycling PCR using sequence-specific probes. The buffer includes proprietary Q-Bond Molecule, which allows short cycling times on standard cyclers and on fast cyclers with rapid ramping rates. The fast-cycling procedure increases throughput by reducing PCR run times by up to 40% (see figure "Genotyping workflow"). 

Convenient kit format

The kit is provided in a ready-to-use, preoptimized master mix format for greater convenience. Use of a master mix saves time, simplifies handling for reaction setup, and increases reproducibility by eliminating many possible sources of pipetting errors and contamination —  pipetting steps are minimized and tedious calculations are eliminated. HotStarTaq Plus DNA Polymerase (included in the master mix) is activated by a 5-minute, 95°C incubation step, which can easily be incorporated into existing thermal cycling programs. Room-temperature reaction setup using the master mix is fast and easy.

Suitable instruments

The Type-it Fast SNP Probe PCR Kit is optimized for amplification of SNP PCR assays on any suitable standard and fast ramping cycler compatible with optical PCR plates that fit into real-time PCR instruments used for fluorescence plate read analysis (see table).

Suitable instruments
Cyclers Model
Real-time PCR cyclers QIAGEN: Rotor-Gene Q
ABI PRISM 7900 (all series) 
ABI StepOne and StepOne Plus
Applied Biosystems 7500 (all series)
Applied Biosystems ViiA 7
Bio-Rad: iCycler iQ
Bio-Rad: CFX series
Roche: LightCycler 480
Roche: LightCycler Nano
Agilent: Mx3000P and Mx3005P
Standard cyclers All cyclers in standard-ramping modes (e.g., GeneAmp 9700)
All cyclers in fast-ramping modes (e.g., GeneAmp 9800)
应用

The Type-it Fast SNP Probe PCR Kit is used to detect SNPs using TaqMan® MGB probes and can be used in various fields of research, including:

  • Typing of disease loci
  • Biomarker discovery
特点
参数
应用 Probe-based SNP Genotyping
酶活 5'-> 3' Exonuclease activity
预混液 Yes
产品使用 Functionally validated and developed for reliable SNP Genotyping
反应类型 PCR amplification
real-time或终点法PCR Both
样本/目标类型 Genomic DNA
序列特异性探针 TaqMan® Genotyping Assays, TaqMan® or TaqMan MGB® probes
有/无ROX ROX included in Master Mix
有/无热启动酶 With

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产品介绍与指南
3
图片
Maximal Success in Automated Allele Calling
成功的自动化等位基因信号分析。
使用6个不同的TaqMan MGB-based SNP基因分型实验和来自60个样本的1 ng基因组DNA对一组不同的DNA进行平均自动化等位基因信号率分析。在384孔板中以5 μl反应体系对指定产物进行PCR。在Applied Biosystems 7900HT仪器上进行等位基因区分读板。Type-it Fast SNP Probe PCR Kit可持续获得最高的信号率和最低的错误率。
Highly Specific Probe Binding due to the Unique SNP Genotyping Buffer System
高度特异性的探针结合。
由于错配探针的非特异性结合,来自其他供应商的用于SNP基因分型的缓冲液经常不能很好分离等位基因簇。Type-it Fast SNP Probe Buffer System通过缩小熔化温度范围,可提高分辨率,减少错配探针的非特异性结合。可对等位基因簇进行广泛的和明确的分离。
Genotyping Workflow Using the Type-it Fast SNP Probe PCR Kit
基因分型工作流程。
Type-it Fast SNP Probe PCR Master Mix含有高度特异性的HotStarTaq Plus Polymerase,可在室温下快速、简单地构建反应体系,即使模板量很少也能确保高度特异的扩增。
Reliable SNP Genotyping even when Using Small Template Amounts
使用少量模板进行可靠的SNP基因分型。
使用[A] Type-it Fast SNP Probe PCR Kit或[B]来自供应商A的基因分型预混液对80个不同基因组DNA(每5 μl反应有10 ng或1 ng)进行等位基因区分。即使使用少量模板,Type-it Fast SNP Probe PCR Master Mix的表现都优于来自供应商A的预混液,获得紧密的等位基因和可靠的结果。使用TaqMan SNP基因分型实验对rs 951134(aryl-hydrocarbon receptor repressor gene)和2个无模板对照(NTCs)进行PCR,在Applied Biosystems 7900HT仪器上进行分析。黑色:NTCs;蓝色:T等位基因纯合子(FAM荧光);绿色:杂合子样本;红色:A等位基因纯合子(VIC荧光)。数据以98%信度进行分析。
Mechanism of Fast Cycling during Annealing
退火过程中快速循环的机制。
[A]专门的PCR缓冲液含有Q-Bond,可以加速退火步骤,将时间减少到5秒。Q-Bond提高Taq DNA聚合酶对短的单链DNA片段的亲和力,减少了引物退火所需时间。[B]没有Q-Bond时,引物和聚合酶依次结合到模板上,增加了退火步骤的持续时间。