计算溶液所需的质量、体积或浓度。
Anstart Taq II DNA Polymerase
有货
为什么选择此级别
EnzymoPure™ 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
热激条件：95℃，2.5min
外切酶活性：5’→3’
防污染体系搭配：√
ARMS：√
SNP：√
荧光定量PCR：√
额外添加AT载体连接：√
推荐项目：定性PCR、定量PCR
产品说明
Anstart Taq II DNA 聚合酶是抗 Taq单克隆抗体和 Taq II DNA 聚合酶的混合制品。是在原Anstart Taq DNA Polymerase (MD006)产品的升级款，具有更好的稳定性和特异性。高温加热前，抗Taq单克隆抗体与Taq聚合酶结合从而抑制聚合酶的活性，达到抑制低温条件下由引物的非特异性退火或引物二聚体引起的非特异性扩增作用。当扩增反应体系被加热到 95℃ 2min时，聚合酶活性因抗Taq单克隆抗体变性得到恢复，因此无需特殊失活处理，在常规PCR反应条件下即可使用。通过与改良的buffer配合使用，快速激活，能有效增加反应产物的量和提高PCR反应灵敏度和特异性。可应用与热启动PCR，扩增复杂模板，低拷贝靶片段扩增。
产品内容
1. Anstart Taq II DNA Polymerase ( 5 U/μl )
2. 10×Anstart Taq II Buffer (without Mg2+ )
3. 100mM MgCl2
产品用途
热启动法进行PCR 扩增。
使用方法
1.PCR 反应体系的设置
a.溶解并混匀 PCR 反应所需的各种溶液，并放置于冰浴上或冰盒内。建议反应 PCR 液体分装使用，避免反复冻融。
b.参考下表设置 PCR 反应，建议 PCR 反应体系的配置在冰浴或在冰盒上进行：
试剂 | 体积 | 终浓度 |
5×Anstart Taq II Buffer (Mg2+ Plus) | 10μl | 1× |
dNTP（25mM each） | 0.4μl | 0.2mM |
Template DNA | 10μl | — |
Primer I（10μM） | 1μl | 0.2μM |
Primer II（10μM） | 1μl | 0.2μM |
Anstart Taq II DNA Polymerase ( 5 U/μl ) | 0.5μl | — |
超纯水 | Up to 50μl | — |
总体积 | 50μl | — |
*对于不同类型的模板在 50μl 反应体积中推荐用量如下：
哺乳动物基因组 DNA：0.1-1μg
大肠杆菌基因组 DNA：10-100ng
质粒 DNA：0.1-10ng
过多的模板 DNA 容易导致非特异性的 PCR 产物
c.用移液器轻轻吹打混匀或轻微 Vortex 混匀，室温离心数秒，使液体积聚于管底。
d.各设置好的 PCR 反应管置于 PCR 仪上，开始 PCR 反应。
2.PCR 反应参数的设置以扩增 1Kb 目的片段为例
步骤 | 温度 | 时间 | 循环数 |
预变性 | 95℃ | 2.5min | 1 |
变性 | 94℃ | 30s |
25-35 |
退火 | 55℃ | 30s | |
延伸 | 72℃ | 1min | |
最后延伸 | 72℃ | 10min | 1 |
a.PCR 反应的设置需根据模板、引物、PCR 产物的长度和 GC 含量等条件的不同设定不同的 PCR 反应条件包括温度、时间和循环数等。
b.STEP4(延伸)的时间设置需根据 PCR 产物的长度进行设置，通常每 kb 产物的延伸时间为 1min。例如 PCR 产物的长度为 1kb，则延伸时间可以设置为 1min，PCR 产物的长度为 2kb，则延伸时间可以设置为 2min，以此类推。
c. 对于初次进行的 PCR，为尽量确保可以扩增出预期的 PCR 产物，可以把循环数设置为 35。对于需进行半定量或定量的 PCR 反应循环数一定要进行适当优化，使 PCR 反应达到平台期。
Heat shock condition: 95 ℃, 2.5min
Externase activity: 5 '→ 3'
Matching of pollution prevention system: √
ARMS: √
SNP: √
Fluorescence quantitative PCR: √
Additional AT carrier connection added: √
Recommended items: qualitative PCR, quantitative PCR
Product manual
Anstart Taq II DNA polymerase is a mixed product of anti-Taq monoclonal antibody and Taq II DNA polymerase. It is an upgraded version of the original Anstart Taq DNA Polymerase (MD006) product, with better stability and specificity. Before heating at high temperature, the anti-Taq monoclonal antibody binds to Taq polymerase to inhibit the activity of the polymerase, thereby inhibiting non-specific annealing of primers or non-specific amplification caused by primer dimers under low temperature conditions. When the amplification reaction system is heated to 95°C for 2 minutes, the polymerase activity is restored due to the denaturation of the anti-Taq monoclonal antibody, so there is no need for special inactivation treatment, and it can be used under conventional PCR reaction conditions. By using it in conjunction with an improved buffer, rapid activation can effectively increase the amount of reaction products and improve the sensitivity and specificity of the PCR reaction. It can be applied to hot-start PCR to amplify complex templates and low-copy target fragments.
Product content
1. Anstart Taq II DNA Polymerase ( 5 U/μl )
2. 10×Anstart Taq II Buffer (without Mg2+ )
3. 100mM MgCl2
Product Usage
The hot start method is used for PCR amplification.
Instructions
1. PCR reaction system settings
a. Dissolve and mix the various solutions required for the PCR reaction, and place them on an ice bath or in an ice box. It is recommended to use aliquots of the reaction PCR liquid to avoid repeated freezing and thawing.
b. Refer to the following table to set up the PCR reaction. It is recommended to configure the PCR reaction system in an ice bath or on an ice box:
Reagent | Volume | Final concentration |
5×Anstart Taq II Buffer (Mg2+ Plus) | 10μl | 1× |
dNTP（25mM each） | 0.4μl | 0.2mM |
Template DNA | 10μl | — |
Primer I（10μM） | 1μl | 0.2μM |
Primer II（10μM） | 1μl | 0.2μM |
Anstart Taq II DNA Polymerase ( 5 U/μl ) | 0.5μl | — |
Ultra-pure water | Up to 50μl | — |
Total capacity | 50μl | — |
The recommended dosage for different types of templates in a 50μl reaction volume is as follows:
Mammalian genomic DNA: 0.1-1μg
Escherichia coli genomic DNA: 10-100ng
Plasmid DNA: 0.1-10ng
Too much template DNA can easily lead to non-specific PCR products
c. Use a pipette to mix gently or gently Vortex to mix, and centrifuge for a few seconds at room temperature to allow the liquid to accumulate at the bottom of the tube.
d. Place each set of PCR reaction tubes on the PCR machine to start the PCR reaction.
2. The setting of PCR reaction parameters takes the amplification of 1Kb target fragment as an example
Step | Temperature | Time | Number of cycles |
Predenaturation | 95℃ | 2.5min | 1 |
Transsexual | 94℃ | 30s |
25-35 |
Annealing | 55℃ | 30s | |
Extend | 72℃ | 1min | |
Last extension | 72℃ | 10min | 1 |
a. PCR reaction settings need to be based on the template, primers, PCR product length and GC content and other conditions to set different PCR reaction conditions, including temperature, time and number of cycles.
b. The time setting of STEP4 (extension) needs to be set according to the length of the PCR product, usually the extension time per kb product is 1 min. For example, if the length of the PCR product is 1kb, the extension time can be set to 1min, and the length of the PCR product is 2kb, then the extension time can be set to 2min, and so on.
c. For the first PCR, in order to ensure that the expected PCR product can be amplified as much as possible, the number of cycles can be set to 35. The number of PCR reaction cycles that need to be semi-quantitative or quantitative must be appropriately optimized to make the PCR reaction reach the plateau.
规格
产品名称
Anstart Taq II DNA Polymerase
别名
启动 Taq II DNA 聚合酶
规格或纯度
EnzymoPure™, 5U/μL
分子类型
酶
产品线
biological_products
储存与运输
浓度
5U/μL
储存条件
-20°C储存
运输条件
超低温运输
技术文档
高级数据
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技术文档和文章
溶液计算器
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