HotStarTaq Plus DNA Polymerase

快速、高特异性扩增，适合多种应用

该产品将于 2023 年 11 月 30 日停产。

立即切换到后续 AllTaq PCR 试剂盒。了解有关过渡的更多信息，并查看详细的常见问答。

访问知识中心

HotStarTaq Plus DNA Polymerase (1000)

Cat. No. / ID: 203605

1000 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl₂

Inquire

HotStarTaq Plus DNA Polymerase 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。

该产品将于 2023 年 11 月 30 日停产。

立即切换到后续 AllTaq PCR 试剂盒。了解有关过渡的更多信息，并查看详细的常见问答。

访问知识中心

特点

高特异性PCR，所需优化次数少
快速激活热启动酶，只需5分钟
即用型PCR上样缓冲液，操作更简单快速

产品详情

该聚合酶具备高特异性、高灵敏度，HotStarTaq DNA Polymerase所需的优化次数少，激活时间为5分钟。可在室温下构建PCR反应体系，且反应产物可直接上样至凝胶，因为新型的CoralLoad PCR缓冲液包含两种凝胶示踪染料。同时提供标准QIAGEN PCR缓冲液，便于操作。此外，一种新型的添加剂Q-Solution可确保高效扩增难扩增的模板（如富含GC的模板）。独特的试剂盒组成和经优化的实验方案使PCR流程更加高效。

绩效

每一批HotStarTaq Plus DNA Polymerase都经过全方位的质量控制测试，包括：严格的PCR特异性和重复性分析，即使低拷贝的靶分子也可扩增。通过检测，HotStarTaq Plus DNA Polymerase优于其它供应者提供的试剂盒，确保高度特异性和在热启动PCR中的卓越表现（参见""、""和表格）。该试剂盒提供的新型PCR缓冲液使得在多种PCR条件下都维持高特异性，无需优化。Q-Solution可进一步优化PCR，且该试剂盒中提供（参见""）。总之，这些组合确保了在多种应用中的特异性扩增（参见""和""）。

热启动模式比较
	HotStarTaq Plus DNA Polymerase	HotStarTaq DNA Polymerase	Supplier A_II提供的热启动酶	供应商R	供应商I （抗体介导）	手动	石蜡屏障
特异性扩增	++	++	+	++	+	+/–	+/–
PCR优化需求	++	++	+/–	+/–	+/–	–	–
易用性	+++	++	++	+	+	–	–
激活速度	++	+	++	++	++	–	–

HotStarTaq Plus DNA Polymerase特性

浓度：5 units/µl
重组酶：是
底物类似物：dNTP、ddNTP、dUTP、biotin-11-dUTP、DIG-11-dUTP、fluorescent-dNTP/ddNTP
延伸速度：72°C条件下2–4 kb/min
酶半衰期：97°C条件下10 min，94°C条件下60 min
扩增效率：≥10⁵倍
5'–>3'外切酶活性：是
额外添加A：是
3'–>5' 外切酶活性：否
核酸酶污染：否
蛋白酶污染：否
RNases污染：否
自启活性：否

查看图表

Highest specificity.

Higher specificity with different primer–template systems.

Amplification of difficult templates.

Effect of hot start on RT-PCR performance.

Highly sensitive single-cell PCR.

原理

HotStarTaq Plus DNA Polymerase具备同HotStarTaq DNA Polymerase一样的卓越表现，激活时间只需5分钟。

HotStarTaq Plus DNA Polymerase，是一种经修饰的QIAGEN Taq DNA Polymerase，以非活性形式提供，常温下没有聚合酶活性。避免PCR构建和循环初始阶段低温条件下非特异性引物的延伸和引物二聚体的形成（参见" Highest specificity"和" Higher specificity with different primer-template systems"）。95°C条件下孵育5分钟即可激活HotStarTaq Plus DNA Polymerase，这个步骤可整合入已有的热循环程序中。

QIAGEN PCR Buffer

QIAGEN PCR Buffer通过提高每个PCR扩增退火过程特异性引物的结合比例，确保PCR每个循环特异性扩增（参见" Increased specificity of primer annealing"）。独特的KCl和(NH₄)₂SO₄平衡组合，使得该缓冲液与传统的PCR缓冲液相比在很宽的温度和Mg²⁺浓度范围内都有严格的引物退火条件和高度的特异性，无需进行耗时的优化。

CoralLoad PCR Buffer

HotStarTaq Plus DNA Polymerase同时提供CoralLoad PCR Buffer，CoralLoad PCR Buffer具备QIAGEN PCR Buffer的所有优点，同时可直接将PCR产物上样到琼脂凝胶上，无需加入凝胶上样缓冲液。CoralLoad PCR Buffer具备同样高的PCR特异性，与常规的QIAGEN PCR Buffer一样可减少优化过程。此外，它包含两种指示染料，橙色染料和红色染料，可方便的判断DNA的迁移距离和优化跑胶时间（参见" CoralLoad PCR Buffer"）。该缓冲液提高移液的可见性，并可直接将PCR产物上样至凝胶，提高便利性。

Q-Solution

该产品同时提供Q-Solution，一种新型的PCR添加剂，通过修饰DNA熔解行为促进难扩增模板的扩增。这种独特的试剂可进一步优化由模板引起的表现不理想的PCR，如过多的二级结构和富含GC的模板（参见" Amplification of difficult templates"）。不同于DMSO和其他PCR添加剂，Q-Solution的浓度是确定的，没有毒性，并且保证PCR的纯度。而且PCR中加入Q-Solution不会影响PCR的准确性。

查看图表

Highest specificity.

Higher specificity with different primer–template systems.

Effect of hot start on RT-PCR performance.

CoralLoad PCR Buffer.

Amplification of difficult templates.

程序

HotStarTaq Plus DNA Polymerase提供高效、经优化的实验方案，用于快速、方便的构建PCR反应体系。95°C条件下孵育5分钟激活HotStarTaq DNA Polymerase，可以整合入已有的热循环程序。反应可在室温下建立，更加方便和易于使用（参见""）。该试剂盒同时提供CoralLoad PCR Buffer，提高移液的可见性，并可直接将PCR产物上样至凝胶，提高便利性。

查看图表

HotStarTaq Plus procedure.

应用

HotStarTaq Plus DNA Polymerase适合多种应用，包括各种富有挑战性的研究，如各种扩增应用：

复杂的基因模板
复杂的cDNA模板（如RT-PCR）
低拷贝的靶分子（如单细胞PCR）
多重引物对反应

辅助数据和图表

CoralLoad PCR Buffer.

Increased specificity of primer annealing.

Specifications

Features	Specifications
Applications	PCR, RT-PCR, Complex genomic templates, very low-copy targets
With/without hotstart	With hotstart
Sample/target type	Genomic DNA and cDNA
Enzyme activity	5' -> 3' exonuclease activity
Reaction type	PCR amplification
Single or multiplex	Single
Mastermix	No
Real-time or endpoint	Endpoint

资源

Second edition — innovative tools

PCR 実験における重要項目と新技術

Addressing critical factors and new solutions

For highly specific hot-start PCR without optimization

Download Safety Data Sheets for QIAGEN product components.

Publications

The putatively functional Mkrn1-p1 pseudogene is neither expressed nor imprinted, nor does it regulate its source gene in trans.

Gray TA; Wilson A; Fortin PJ; Nicholls RD;

Proc Natl Acad Sci U S A; 2006; 103 (32):12039-44 2006 Aug 1 PMID:16882727

Use of a short fragment of the C-terminal E gene for detection and characterization of two new lineages of dengue virus 1 in India.

Domingo C; Palacios G; Jabado O; Reyes N; Niedrig M; Gascón J; Cabrerizo M; Lipkin WI; Tenorio A;

J Clin Microbiol; 2006; 44 (4):1519-29 2006 Apr PMID:16597885

FAQ

No. HotStarTaq Plus DNA Polymerase is supplied with conventional QIAGEN PCR Buffer and CoralLoad PCR Buffer in separate vials. Both buffers minimize nonspecific amplification products, primer–dimers, and background.

CoralLoad PCR Buffer has all of the advantages of QIAGEN PCR Buffer but can also be used to directly load the PCR reaction onto an agarose gel without the need to add a gel loading buffer.

FAQ ID -1052

PCR products that will be cloned using the QIAGEN PCR Cloning Kit should be generated using a thermostable DNA Polymerase without proofreading activity, such as Taq DNA Polymerase. Such polymerases attach a single A overhang to their reaction products, which can hybridize to the U overhang of the pDrive Cloning Vector. For efficient addition of an A overhang during the PCR procedure, we recommend a final extension step for 10 min at 72°C as described in the standard protocols of the Taq PCR- and HotStarTaq PCR handbook.

FAQ ID -165

Yes, please follow the Supplementary Protocol 'Polyacrylamide_gel_analysis_of_oligonucleotides' (PCR03).

FAQ ID -961

Not necessarily. In a lot of cases, the uniquely formulated PCR Buffer provided in the HotStarTag Plus DNA Polymerase, HotStar HiFidelity Polymerase, Taq DNA Polymerase, HotStarTaq DNA Polymerase, and QIAGEN Multiplex PCR Kits provides optimal amplification of specific PCR products. The usefulness of Q-Solution needs to be determined empirically for each primer/template setup, by running parallel PCR reactions with and without Q-Solution under the same cycling conditions.

Q-Solution changes the melting behavior of DNA and will often improve a suboptimal PCR caused by templates that have a high degree of secondary structure or high GC-contents. For more details on the effects of Q-Solution on PCR amplification, please see the Q-Solution sections of the HotStarTaq Plus DNA Polymerase, HotStar HiFidelity Polymerase, Taq DNA Polymerase, HotStarTaq DNA Polymerase, and the QIAGEN Multiplex PCR Handbooks.

FAQ ID -380

Yes. Do not increase the activation time for HotStarTaq Plus DNA Polymerase beyond the recommended 5 minutes, as PCR product yield may be reduced and less specific amplification may result. Please follow the instructions in the HotStarTaq Plus PCR Handbook closely, and use the cycling parameters presented in the handbook to ensure optimal results.

FAQ ID -1050

CoralLoad gel tracking dye contained in Taq, HotStarTaq, TopTaq DNA Polymerase and TopTaq Master Mix Kits separates into 2 fragment-size dependent colors (orange and red) when loaded onto an agarose gel. Sigma Red buffer only has one color which is harder to visualize.

FAQ ID -1644

The activation time for HotStarTaq Plus DNA Polymerase used in the QuantiFast SYBR Green PCR Kits is longer than that for QuantiFast Probe PCR Kits. This is due to differences in buffer composition. Buffer components such as salts and additives influence the time required for enzyme activation.

FAQ ID -1449

Yes. Separated dyes are visible on an agarose gel down to a volume of 2 µl of PCR product generated with HotStarTaq Plus DNA Polymerase and CoralLoad PCR buffer.

FAQ ID -1051

In QIAGEN labs, we have amplified PCR products up to 5 kb from complex genomic DNA, and up to 10 kb from less complex lambda phage DNA with the HotStar HiFidelity Polymerase Kit, following standard protocols in the HotStar HiFidelity PCR Handbook.

For targets larger than 5 kb of complex genomic DNA, and larger than 10 kb of less complex DNA, we recommend to follow the protocol 'Amplification of Long PCR Products' in the HotStar HiFidelity PCR Handbook. The protocol uses a mixture of HotStar HiFidelity DNA Polymerase and Taq, or HotStar Taq Plus DNA Polymerase, and allows much longer fragments to be generated. In-house we have tested fragments up to 13 kb from complex genomic DNA or up to 30 kb from less complex lambda phage DNA using this protocol.

FAQ ID -1047