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QIAGEN Fast Cycling PCR Kit

用于超快速和特异性PCR,适合各种PCR仪

S_1312_AppD_HST0442
This product is in transition
A successor version of this kit is now available. Learn more about the transition and see detailed FAQs.

QIAGEN Fast Cycling PCR Kit (200)

Cat. No. / ID:   203743

For 200 x 20 μl reactions: 2 x 1 ml QIAGEN Fast Cycling PCR Master Mix, 10x CoralLoad Dye, 5x Q-Solution, RNase-Free Water
AU$296.00
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Preparations
200
1000
QIAGEN Fast Cycling PCR Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。
This product is in transition
A successor version of this kit is now available. Learn more about the transition and see detailed FAQs.

Features

  • 热启动PCR扩增,少于15分钟(35个循环)
  • 适合各种PCR仪器
  • 即用型预混液含有HotStarTaq Plus DNA Polymerase
  • 可直接用于电泳上样的PCR缓冲液,操作简单快速
  • 无需为快速PCR重新设计引物

Product Details

快速循环PCR预混液包含HotStarTaq Plus DNA Polymerase适合高特异性高灵敏度的PCR反应,以及独特的缓冲液体系用于极短时间的变性、退火和延伸步骤。专有的PCR缓冲体系明显缩短了聚合酶、引物和模板复合体形成所需的时间,因此将总的PCR循环时间从1.5小时左右缩短到20分钟。Optional CoraLoad Dye(同样包含在该试剂盒中)包括胶示踪染料,使分析更加简单。此外,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。

Performance

QIAGEN Fast Cycling PCR Kit拥有高性能的HotStarTaq Plus DNA Polymerase以及在各种热循环仪上显著缩短的PCR循环时间。不同于其他常会致扩增失败的快速循环试剂,HotStarTaq Plus DNA Polymerase与QIAGEN Fast Cycling PCR Buffer结合使用,促进了特异性引物-模板退火并显著降低非特异性退火(参见" Specific and reliable results")。可在每一个PCR循环中减少非特异性产物的扩增、以及引物二聚体的形成。使用创新的专有PCR缓冲液制定方案节约时间高达75%。PCR循环时间可以从超过1小时(35个循环)减少到少于20分钟。快速升温的PCR仪可以在30分钟内实现总PCR(参见" Time savings of up to 75%")。可用Q-Solution,包含在该试剂盒中的新型辅助剂,优化PCR。
See figures

Principle

QIAGEN Fast Cycling PCR Kit以混合液规格提供,包含在每个循环仪上进行快速特异性扩增需要的所有组分(参见下表)。可有效扩增长达3.5 kb的基因组DNA和cDNA的PCR产物,包含复杂的模板。HotStarTaq Plus DNA Polymerase确保了高度特异性。使用新型的专利PCR缓冲液制定方案可显著节省时间高达75%,有效降低了形成聚合酶、引物和复杂模板。这能够让聚合酶在PCR退火阶段延伸引物,因此能有效缩短每个PCR循环中的退火过程。PCR循环时间可从超过1小时(35个循环)降低到少于20分钟(参见" Significant time savings")。快速升降温的PCR仪可将整个PCR时间控制在30分钟内(参见"Time savings of up to 75%"))。

可靠的快速循环PCR所需组分
试剂盒组分 特征
HotStarTaq Plus DNA Polymerase 高特异性产物(多达3.5 kb)
高特异性 
室温设置
Fast Cycling PCR Buffer(包含Q-Bond Molecule) 高速 
需要最少的优化
CoralLoad Dye 可立即用凝胶加载PCR产物 
优化的可视移液器
Q-Solution 方便扩增的高GC模板
QIAGEN Fast Cycling PCR Master Mix 方便快速的反应设置

高特异性和灵敏度

HotStarTaq Plus DNA Polymerase以混合液规格提供,确保了良好的热启动PCR性能。所提供的HotStarTaq Plus DNA Polymerase不具活性,在室温下没有聚合酶活性。因此可以防止较低温度时形成错配产物和引物二聚体(参见"Specific and reliable results"),同时使常温下PCR体系的构建更加方便。 HotStarTaq Plus DNA Polymerase在95°C下,由短时的5分钟诱导激活,可以较易地纳入到各种现有的PCR仪程序中。

独特的快速循环PCR缓冲液

新型的Fast Cycling PCR Buffer优化了特异性PCR产物的扩增,并显著地减少了循环时间。基于原始的QIAGEN PCR Buffer,这种新的制备方案在每个PCR循环的短时退火步骤中,确保了特异性-非特异性引物结合的高比率。由于KCl和(NH4)2SO4独特的平衡结合,相对于传统的PCR缓冲液,该PCR缓冲液提供严格的引物退火条件,具有更广泛的退火温度和Mg2+浓度。使用这种缓冲体系可在宽泛的温度范围和Mg2+浓度下获得特异性扩增,因此可有效减少PCR的优化。

创新的快速循环缓冲液包含新型的Q-Bond Molecule。这种分子可以极大地提高DNA聚合酶和单链DNA的结合能力,因此有助于将退火时间减少到仅需5秒(参见"Mechanism of fast cycling during annealing")。独特的缓冲体系配方和经优化的DNA聚合酶浓度都极大地降低了变性和延伸时间。Fast Cycling PCR Buffer被开发,从而无需对每个引物-模板体系进行优化,节省了时间和经费。使用这种缓冲体系可在宽泛的温度范围和Mg2+浓度下获得特异性扩增,而省去了耗时的优化过程。

CoralLoad染料

QIAGEN Fast Cycling PCR Kit包含新型的CoralLoad Fast Cycling Dye。这种染料可以将PCR产物直接上样到琼脂糖凝胶中而无需加入凝胶上样缓冲液(参见"Ready-to-load PCR reactions")。CoralLoad Fast Cycling Dye包含2种染料:一种橙黄色染料和一种红色的染料,有助于估计DNA迁移距离并优化凝胶电泳时间。该染料提供了优化的移液器可视性并能直接方便地将PCR产物上样到凝胶上。

Q-Solution

QIAGEN Fast Cycling PCR Kit还包含一种新型的PCR辅助剂Q-Solution,可通过改变DNA的熔解过程,来辅助困难模板的扩增。这种独特的试剂可以改善由于大量的二级结构或高GC含量的模版所导致的不理想扩增。与常用的其他PCR辅助剂,如DMSO不同,Q-Solution只有一个工作浓度,并且保证无毒和高纯度。

See figures

Procedure

QIAGEN Fast Cycling PCR Kit为方便用户使用提供方便的混合液规格。HotStarTaq Plus DNA Polymerase在95°C下经5分钟诱导激活,此步骤可方便的纳入现有的热循环方案中。使用混合液的室温反应设置快速简易。将移液步骤最小化,减少了失误和污染的可能性,确保增加了通量和可重复性。该试剂盒包含用于快速简单PCR设置的精简优化实验方案。同样有该试剂盒提供的CoralLoad Dye增加了移液器的可视性,并确保可直接方便地将PCR产物上样到凝胶样上。

PCR循环时间可从超过1小时(35个循环)减少至少于20分钟(参见"Significant time savings"和下表"用于不同片段长度的PCR循环时间")。快速升温PCR仪可仅在30分钟内得到总PCR(参见Time savings of up to 75%")。The time savings afforded by the 由于QIAGEN Fast Cycling PCR Kit可节省时间,使其高度是用于通量增加的PCR反应,并且无需购买额外的昂贵实验设备。

节省大量时间

QIAGEN Fast Cycling PCR Kit通过使用现有的实验设备增加PCR通量。该试剂盒可用于各种PCR仪,包括无特异性缺失的快速升温PCR仪(参见"Time savings of up to 75% ")。此外,不同于其他PCR试剂,使用该试剂无需调整或改变引物或退火温度。每个试剂盒组分经精心设计以提供方便可靠的快速OCR(参见" Easy and convenient procedure"和下表)。

不同片段长度的PCR循环时间
片段长度(bp) QIAGEN Fast Cycling操作流程(min) 标准循环操作流程(min) 节省时间(%)
200 15 68 78
500 20 68 71
1000 29 85 66
3000 63 155 59
See figures

Applications

QIAGEN Fast Cycling PCR Kit可用于各种PCR仪,能够增加PCR通量,并适用于高灵敏度和高特异性的PCR和常规PCR,以及以复杂基因组DNA和cDNA作为模板的PCR。该试剂盒高度适用于标准和快速PCR仪。

Supporting data and figures

Specifications

FeaturesSpecifications
Real-time or endpointEndpoint
ApplicationsPCR, RT-PCR, Complex genomic templates, very low copy targets
Sample/target typeGenomic DNA and cDNA
With/without hotstartWith hotstart
Reaction typePCR amplification
MastermixYes
Single or multiplexSingle
Enzyme activity5' -> 3' exonuclease activity

Resources

试剂盒操作手册 (1)
For fast and specific PCR on any thermal cycler  
Safety Data Sheets (3)
Download Safety Data Sheets for QIAGEN product components.
产品介绍与指南 (2)
Second edition — innovative tools
Addressing critical factors and new solutions
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Brochures & Guides (2)
Second edition — innovative tools
Addressing critical factors and new solutions
Kit Handbooks (1)
For fast and specific PCR on any thermal cycler  

FAQ

Do you have a protocol for polyacrylamide gel analysis of oligonucleotides?
Yes, please follow the Supplementary Protocol 'Polyacrylamide_gel_analysis_of_oligonucleotides' (PCR03).
FAQ ID -961
How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling?

Our unique multiplex PCR buffer system with ammonium and potassium ions and Factor MP has been further optimized in QuantiFast and Rotor-Gene Kits. We have also discovered Q-Bond, a buffer component which supports the rapid formation of the polymerase–primer–template complex, leading to reduced annealing times.

FAQ ID -1430
What is Q-Bond used in the QIAGEN Fast Cycling PCR and QuantiFast Kits?

The Q-Bond Molecule, present in the PCR Buffer of the QIAGEN Fast Cycling PCR Kit and the QuantiFast Kits, dramatically increases the binding affinity of DNA polymerase to single-stranded DNA, thereby facilitating the reduction of annealing time to just a few seconds. It is a non-protein PCR component. Unfortunately, all further information on this molecule is proprietary.

FAQ ID -1554
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