高度可靠的终点法PCR,无可比拟的方便性

  • 易于使用的预混液,操作步骤最少化
  • 冷藏储存节省了解冻时间
  • 可靠的高产率PCR
  • 方便的室温操作
  • 优化的操作程序适合所有PCR实验

TopTaq Master Mix Kit结合了TopTaq DNA Polymerase和即用型预混液的所有优势,简单易用,包含TopTaq DNA Polymerase,将优化需求最小化的TopTaq PCR Buffer和dNTPs。酶保存缓冲液中有独特的TopTaq稳定剂,使得TopTaq Master Mix可保存在4°C,并在室温下进行反应,无需耗时的解冻试剂。所有的成分都含在即用型混合液中,减少了移液步骤,降低了污染的风险。TopTaq Master Mix即使在25°C或4°C下存放很长一段时间,还是可以获得高产率的PCR产物。

Product Product no. Cat. no. List price:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
Product Cat. no. List price:
TopTaq Master Mix Kit (250)
For 200 x 50 µl reactions: 2x TopTaq Master Mix containing 250 units of TopTaq DNA Polymerase in total, 10x CoralLoad Concentrate, and RNase-Free Water
200403
Inquire
The TopTaq Master Mix Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
  • Main Image Navi
Reliable high-yield PCR, independent of storage conditions.|Convenient CoralLoad Concentrate.|TopTaq procedure.|High yields of PCR product without optimization|Increased specificity of primer annealing.|Stabilization at 4°C.|
PCR was performed using TopTaq Master Mix stored at –20°C, 4°C, and 25°C for 4 months. Two different genes from human genomic DNA (prp, 750 bp and HugL, 1200 bp) were amplified according to the standard optimized protocol (duplicates shown). The results show high yields of specific PCR product. [A] Gel image produced by the QIAxcel System. Markers: GelPilot 1 kb Plus Marker and QX Alignment Marker 15 bp/10 kb. [B] The QIAxcel System fully automates sensitive, high-resolution capillary electrophoresis of up to 96 samples per run.|[A] CoralLoad Concentrate contains two gel-tracking dyes, enabling immediate gel loading of PCR products for [B] easy visualization of DNA migration.|Due to the unique proprietary TopTaq Stabilizer contained in the enzyme storage buffer, TopTaq DNA Polymerase is the first PCR kit that can be stored at 4°C. TopTaq Stabilizer binds to TopTaq DNA Polymerase, preventing polymerase inactivation. Template DNA and primers are also prevented from binding to the polymerase at low temperatures. During the initial denaturation step, the TopTaq Stabilizer freely dissociates from the polymerase without compromising polymerase activity. This added convenience allows immediate reaction setup without the use of ice — saving the time usually required for reagent thawing.|[A] TopTaq DNA Polymerase was used to amplify 14 different PCR products from mammalian genomic DNA ranging in size from 100 bp to 2 kb according to the standard preoptimized protocol and using identical cycling conditions. [B] The same four targets (shown in lanes 11–14) were amplified under identical cycling conditions using DNA polymerases from the indicated suppliers. TopTaq DNA Polymerase (QIAGEN) resulted in high yields of specific PCR products, regardless of fragment size.|Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K+ binds to the phosphate groups (P) on the DNA backbone, stabilizing the annealing of the primers to the template. NH4+, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing the weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains a high ratio of specific-to-nonspecific primer-template binding over a wide temperature range.|[A] TopTaq DNA Polymerase is stable at 4°C or room temperature due to the presence of low-molecular-weight molecules (TopTaq Stabilizer) on the surface of the polymerase. [B] At higher temperatures, the TopTaq Stabilizer completely dissociates from the polymerase without compromising polymerase activity.|
Performance
TopTaq Master Mix Kit用于标准的和特有的终点式PCR应用,无需优化。TopTaq Master Mix即使在25°C或4°C下保存4个月还是可以获得高产率的PCR产物,说明TopTaq Master Mix能够提供非常稳定和可靠的扩增体系(参见"Reliable high-yield PCR")。TopTaq稳定剂与TopTaq DNA Polymerase结合,防止聚合酶在长期储存过程中灭活。预混液中含有独特的PCR,具有均衡的钾和铵离子,改进特异性引物-模板的退火条件,并同时降低非特异性退火条件。TopTaq DNA Polymerase比其他供应商测试的DNA聚合酶具有更好的性能,确保了即使在使用较低的模板量,也可获得高产量的特异性产物和单独的片段大小(参见"High PCR product yields without optimization”)。
Principle

由TopTaq Master Mix Kit研发用于提供前所未有的应用便捷性和高度可靠的终点式PCR。在研发TopTaq PCR体系之前,所有PCR酶都需要在–20°C下储存。因为在酶保存缓冲液中含有独特的专有TopTaq稳定剂,TopTaq Master Mix Kit是首款可在4°C下储存的试剂盒。该即用型预混液规格提供了更大的便利。由于TopTaq DNA Polymerase的使用,甚至在扩增不同大小的片段时,也可获得高产量的特异性产物。(参见"High PCR product yields without optimization")。

TopTaq PCR Buffer

新型的TopTaq PCR Buffer有助于扩增特异性PCR产物。在每个PCR循环的退火步骤中,相对非特异性引物结合,该缓冲液大大增加了特异性引物结合的比率。由于KCl和(NH4)2SO4独特均衡的结合,相比传统的PCR缓冲液,该PCR缓冲液提供了更广泛的温度和Mg2+浓度,以及严格的引物-退火条件(参见"Increased specificity of primer annealing")。通过调节退火温度或Mg2+浓度可显著减少优化PCR反应的需求,有时不需要优化过程。

TopTaq稳定剂

TopTaq与TopTaq DNA Polymerase结合,防止在长期储存过程中聚合酶灭活(参见"Stabilization at 4°C"),同时防止模板DNA和引物在低温下与聚合酶结合。在初始的降解步骤中,TopTaq稳定剂能从聚合酶中分离而不影响聚合酶的活性。

CoralLoad Concentrate

为了更方便使用,成套提供TopTaq Master Mix Kit和CoralLoad Concentrate。CoralLoad Concentrate包含两种胶示踪染料。该独特的试剂提高了移液可视化,有助于去除DNA迁移距离并优化琼脂糖凝胶跑样时间。使用CoralLoad Concentrate时,可将PCR产物直接上样到琼脂糖凝胶上而不需要事先另外加缓冲液(参见"Convenient CoralLoad Concentrate")。

Procedure
TopTaq Master Mix Kit结合了独特的TopTaq DNA Polymerase和即用型预混液的所有优势。由于TopTaq Master Mix Kit,无需单独的移液部分组件,并能最大限度地减少污染风险和移液可视化,使PCR反应体系构建快速简单。在初始的降解步骤中,TopTaq稳定剂能从聚合酶中分离而不影响聚合酶活性。这增加了便利,并能即可进行反映设置,无需冰上工作,节省了解冻试剂所需的时间(参见"TopTaq procedure")。
可靠的高产量PCR,无需优化过程

经优化的实验方案适用于所有PCR检测,去除了个别引物-模板体系中漫长的实验参数优化需要,能节省大量的时间和成本。TopTaq Master Mix即使在25°C或4°C下保存4个月还是可以获得高产率特异性PCR产物(参见"Reliable high-yield PCR")。

Applications

TopTaq Master Mix Kit高度适用于所有终点法PCR应用。

Feature
Specifications
Applications PCR, RT-PCR, DNA Fingerprinting
dNTP's included Yes (in Master Mix)
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 Single
Storage conditions Fridge storage and room temperature setup
With/without hotstart Without Hotstart

You are not authorized to download the resource

Kit Handbooks
1
For standard and specialized end-point PCR applications without the need for optimization
Show details