Type-it Fast SNP Probe PCR Kit

应用TaqMan或TaqMan MGB探针进行准确可靠的SNP基因分型

  • 经TaqMan SNP Genotyping Assays验证
  • 自动等位基因分型,致密的荧光光束
  • 适用于难以分型的SNP位点和少量模板
  • 快速循环可节约多达40%的时间
Type-it Fast SNP Probe PCR Kit基于高度特异性的HotStarTaq Plus DNA Polymerase和新研发的缓冲液体系,可获得可靠、清晰的等位基因分型。预混液中所有组分的配合可提高探针结合的准确性和特异性。Type-it Fast SNP Probe PCR Kit经TaqMan SNP Genotyping Assays验证,能获得可重复的SNP基因分型结果,也可用于难处理的SNP位点(如富含GC)或起始量低的模板。
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Type-it Fast SNP Probe PCR Kit (100)
For 100 x 25 μl reactions: 2x Type-it Fast SNP Probe PCR Master Mix, 5x Q-Solution, RNase-Free Water
206042
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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
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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
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The Type-it Fast SNP Probe 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 automated allele calling.|Highly specific probe binding.|Genotyping workflow.|Reliable SNP genotyping with small amounts of template.|Mechanism of fast cycling during annealing.|
Average automated allele call rates for a panel of different DNAs were analyzed using 6 different TaqMan® MGB-based SNP genotyping assays and 1 ng of genomic DNA from each of the 60 samples. PCR was performed with the indicated products in 384-well plates in a 5 μl reaction volume. Allelic discrimination plate read was performed on an Applied Biosystems 7900HT instrument. The Type-it Fast SNP Probe PCR Kit consistently resulted in the highest call rates and the lowest error rates.|Buffer systems from other suppliers used for SNP genotyping often show poorer separation of allele clusters due to nonspecific binding of the mismatch probe. The Type-it Fast SNP Probe Buffer System provides increased discrimination by narrowing the probe melting temperature window, thereby reducing nonspecific binding of the mismatch probe. Wide and clear separation of allele clusters is obtained.|The Type-it Fast SNP Probe PCR Master Mix consists of highly specific HotStarTaq Plus Polymerase, which enables fast and easy reaction setup at room temperature and ensures highly specific amplification — even with low template amounts.|Allelic discrimination was performed with a panel of 80 different genomic DNAs (10 ng or 1 ng each in 5 μl reactions) using either the [A] Type-it Fast SNP Probe PCR Kit or [B] a genotyping master mix from Supplier A. Even when using low amounts of template, the Type-it Fast SNP Probe PCR Master Mix outperformed the master mix from Supplier A and provided tight clustering of alleles and reliable results. PCR was performed with a TaqMan® SNP genotyping assay for rs 951134 (aryl-hydrocarbon receptor repressor gene) and 2 no template controls (NTCs) and analyzed on an Applied Biosystems 7900HT instrument. Black: NTCs; blue: homozygous for T allele (FAM fluorescence); green: heterozygous samples; red: homozygous for A allele (VIC fluorescence). Data was analyzed with 98% confidence.|[A] The proprietary PCR buffer contains Q-Bond, a molecule that facilitates the reduction of the annealing step to 5 seconds. Q-Bond increases the affinity of Taq DNA polymerases for short single-stranded DNA fragments, reducing the time needed for successful primer annealing. [B] Without Q-Bond, the primer and polymerase bind sequentially to the template, increasing the duration of the annealing step.|
Performance

Type-it Fast SNP Probe PCR Kit确保持续获得高度准确的SNP基因分型。

等位基因特异性的探针可进行高度严谨和特异的结合  

Type-it Fast SNP Genotyping PCR Master Mix基于高度特异性的HotStarTaq Plus DNA Polymerase和新研发的SNP基因分型PCR缓冲液体系,可进行高度特异性的探针结合和持续强的荧光信号。与其他商业化SNP基因分型预混液相比,可获得更广泛、更清晰的等位基因群分离(参见"Highly specific probe binding")。

低模板量也能获得高信号率

对于有挑战性的靶点或难处理的SNP位点,Type-it Fast SNP Probe PCR Kit提供可靠的SNP基因分型,可完全分开等位基因并使相同的等位基因紧密成群,产生较高的信号率(参见"Successful automated allele calling")。Type-it Fast SNP Probe PCR Kit优于其他供应商的试剂盒,保证等位基因紧密成群,即使1 ng的DNA模板(参见 "Reliable SNP genotyping with small amounts of template")。

Principle

Type-it Fast SNP Probe PCR Kit采用方便的预混液规格,含有高度特异性的HotStarTaq Plus Polymerase和独特的SNP基因分型缓冲液体系,可快速、高效的扩增。独特的试剂盒组分可使等位基因紧密成群,完全分开确保高信号率,获得可重复的、准确的结果(见表)。

高特异性和灵敏度

预混液中提供的HotStarTaq Plus DNA Polymerase确保高特异性扩增,即使处理低模板量。HotStarTaq Plus DNA Polymerase处于非活化状态,在环境温度下没有聚合酶活性。这避免了在低温下引物错搭产物和引物二聚体的形成。

独特的缓冲液体系

预混液中含有的Type-it Fast SNP PCR Buffer专用于通过序列特异性5'核酸酶探针进行快速循环的SNP基因分型。Type-it Fast SNP PCR Buffer的独特组分确保非常严格和特异性的等位基因特异性探针的结合。改变探针的熔解行为可形成更窄的探针熔解温度窗口。基于原始的QIAGEN PCR Buffer,这种创新的缓冲液使每个PCR循环的退火步骤都有较高比率的特异性引物结合。KCl和(NH4)2SO4独特比例结合,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。

Type-it Fast SNP PCR缓冲液中的添加剂,如Q-Solution,可提供反应条件,用于难处理基因组区域和SNP位点的扩增。创新的Q-Bond技术(用于快速循环),可更快获得SNP基因分型结果(参见"Mechanism of fast cycling during annealing")。预混液中含有的ROX染料,其浓度使其能在Applied Biosystems的多种仪器上进行SNP基因分型。而且,ROX染料也能在不需要ROX作为阴性参照染料的仪器上使用(见表)。

试剂盒的特征
试剂盒组分 特征
2x Master Mix format* 用于配合TaqMan MGB探针进行SNP基因分型。能与多种循环仪配合使用,用于SNP基因分型
HotStarTaq Plus DNA Polymerase 室温下进行快速、简单的反应体系构建。即使使用少量模板,也能高度特异性扩增   
Type-it Fast SNP Probe PCR Buffer 广泛分离基因群。紧密的等位基因群,高等位基因信号率。探针结合具有更高特异性。使用Q-Bond Molecule,快速循环   
Q-Solution 对于高GC含量的SNP位点,在散点图分析中具有更紧密的集群和更强信号。可进一步提高不理想的等位基因信号
* 包括ROX参照染料、dNTPs和MgCl2。
Procedure

Type-it Fast SNP Probe PCR Kit使用商业化的SNP基因分型检测进行了功能验证,能与TaqMan MGB Probes配合使用,也可配合用户自行设计的探针法检测,可使用TaqMan MGB、TaqMan或其他双标记探针。此试剂盒包括简化的、预优化的操作流程,用于快速、可靠的分析。

快速循环步骤,可直接获得结果

Type-it Fast SNP Probe PCR Master Mix提供的反应条件,配合序列特异性探针进行快速PCR循环。缓冲液包含专有化的Q-Bond Molecule,可在标准循环仪或具有快速升温速率的快速循环仪上缩短循环时间。快速循环步骤可减少至多40%的PCR运行时间,提高通量(参见"Genotyping workflow")。

方便的试剂盒规格

该试剂盒采用即用型、预优化预混液形式,具有更大便利性。使用预混液可节约时间,简化反应体系构建的手工操作,通过消除移液误差和污染的可能来源,提高可重复性,尽量减少移液步骤,无需繁琐的计算。预混液中含有的HotStarTaq Plus DNA Polymerase 95°C,5分钟被激活,可在热循环程序中简单设置。使用预混液进行室温下的反应体系构建快速、简单。

适合的仪器

Type-it Fast SNP Probe PCR Kit经优化,可在多种合适的标准和快速升温的循环仪上进行SNP PCR分析,光学PCR孔板配合real-time PCR仪可用于荧光板阅读分析(见表)。

Suitable instruments
循环仪 模式
Real-time PCR cyclers QIAGEN: Rotor-Gene Q
ABI PRISM 7900 (all series)
Applied Biosystems 7500 (all series)
Applied Biosystems 7300
ABI PRISM 7700
ABI PRISM 7000
ABI StepOne and StepOnePlus
Bio-Rad: iCycler iQ
Roche: LightCycler 480
Stratagene: Mx3000P and Mx3005P
Standard cyclers 采用标准升温模式的所有循环仪(如GeneAmp 9700)
采用快速升温模式的所有循环仪(如GeneAmp 9800)

 

Applications

Type-it Fast SNP Probe PCR Kit应用TaqMan MGB探针检测SNPs,可用于多个研究领域,包括:

  • 疾病基因位点的分型
  • 生物标记物的发现
Feature
Specifications
Applications Probe-based SNP Genotyping
Enzyme activity 5'-> 3' Exonuclease activity
Mastermix Yes
Product use Functionally validated and developed for reliable SNP Genotyping
Reaction type PCR amplification
Real-time or endpoint Both
Sample/target type Genomic DNA
Sequence specific Probe TaqMan® Genotyping Assays, TaqMan® or TaqMan® MGB probes
With or without ROX ROX included in Master Mix
With/without hotstart With

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