计算溶液所需的质量、体积或浓度。
为什么选择此级别
，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,避免反复冻融。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
产品内容
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产品简介
本品是由一款新型高效的快速DNA Polymerase、Mg2+、dNTPs以及PCR稳定剂 和增强剂组成的预混体系，浓度为2×。本产品为独创的新型快速DNA聚合 酶，具有极高的扩增速度与稳定性，延伸速度可达5 s/kb，最短十五分钟完成PCR，长 片段（大于3 kb）或者复杂模板可以采用10-30 s/kb的延伸速度或者较多循环数。独创 的MasterMix配方使整个反应体系非常稳定，同时复杂模板也能得到有效扩增，超过 98%的PCR扩增能一次成功。使用时只需要加入DNA模板和引物，补足水即可反应， 可最大限度地减少人为误差、降低污染和节约时间。 本产品已加入染料（蓝色），反应结束后可直接进行电泳检测。扩增得到的PCR 产物3′端附有一个“A”碱基，因此可直接用于T/A克隆，并适用于无缝克隆试剂盒、T4连接试剂盒和感受态产品。 本产品主要适用于超快PCR、复杂模板、复杂二级结构、对高保真性有要求的基 因克隆等实验和大规模基因检测。
质量控制
经检验无外源核酸酶活性；PCR方法检测无宿主残余DNA；能有效地扩增多种基 因组中的单拷贝基因。
使用方法
以下举例为以人基因组DNA为模板，扩增1 kb的片段的PCR反应体系和反应条 件，实际操作中应根据模板、引物结构和目的片段大小不同进行相应的改进和优化。
PCR反应体系
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注意：引物浓度请以终浓度0.1-1.0 μM作为设定范围的参考。扩增效率不高的情况下，可提高引物的 浓度；发生非特异性反应时，可降低引物浓度，由此优化反应体系。
PCR反应条件
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注意： 1）注意：对于简单模板，预变性时间可控制在30 s-1 min,对于复杂模板如菌液等，预变性时间 可增加至2 min。
优化参数设定
1. 模板DNA量设定：
模板过量可能导致非特异性扩增或smear。50 μl PCR反应体系中模板DNA推荐使 用量如下：
-人基因组DNA 5 ng-500 ng
-大肠杆菌基因组DNA 50 pg-100 ng
-质粒DNA 10 pg-1 ng 1. 30-35 个循环数
2 .引物浓度设定： 引物浓度可设为0.1 μM-1.0 μM之间。引物浓度过低时可能导致扩增产物少。引物 浓度过高会抑制特异性扩增，可能导致非特异性扩增。
3. 退火温度设定： 一般实验中退火温度比扩增引物的熔解温度Tm低5℃，无法得到理想的扩增效率 时可适当降低退火温度；发生非特异性反应时可适当提高退火温度。对于复杂模 板，需要调节退火温度实现高效扩增。
4. 延伸时间设定： 延伸时间应根据所扩增片段大小设定。以下为推荐的延伸时间： 质粒等简单模板：5-15 s/kb； 常规基因组、cDNA模板：10-15 s/kb； 复杂模板、粗提模板：20-30 s/kb； （延伸时间不宜过短应至少在5 s/kb以上，也不宜超过30 s/kb）。
5.
循环数设定：
可根据扩增产物的下游应用设定循环数。如果循环次数太少，扩增量不足；如果
循环次数太多，错配机率会增加，非特异性背景严重。所以在保证产物得率的前
提下应尽量减少循环次数。
Product Content:
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Products Introduction
This product is a premixed system consisting of a new high efficient fast DNA Polymerase, Mg2+, dNTPs, and PCR stabilizers and enhancers at 2× concentration. It is a new rapid DNA polymerase developed by CombiSigma with high amplification speed and stability. The extension speed is up to 5 s/kb, and the PCR can be completed in as little as 15 minutes, while longer fragments (>3 kb) or complex templates can be extended at a speed of 10-30 s/kb or a higher number of cycles. The unique MasterMix formula makes the whole reaction system very stable, while complex templates can be amplified effectively, and more than 98% of PCR amplification can be successful in one run. Simply add the DNA template and primers and top up with water to minimize human error, contamination and time.
The dye (blue) has been added to the product and it is ready for electrophoretic detection at the end of the reaction. The PCR product is amplified with an 'A' base at the 3′ end and can therefore be used directly for T/A cloning and is suitable for use in the CombiVerge Seamless Cloning Kit, T4 Ligation Kit and sensory products.
This product is mainly suitable for ultra-fast PCR, complex templates, complex secondary structures, gene cloning and large-scale genetic testing that requires high fidelity.
quality control
No exogenous nuclease activity was detected; no host residual DNA was detected by PCR; single-copy genes in various genomes could be amplified efficiently.
Usage
The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template, which should be improved and optimized according to the template, primer structure and size of the target fragment in actual operation.
PCR reaction system
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Note: Please use the final concentration of 0.1-1.0 μM as a reference for setting the range of primer concentration. If the amplification efficiency is not high, the primer concentration can be increased; if a non-specific reaction occurs, the primer concentration can be decreased to optimize the reaction system.
PCR reaction conditions
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Note: 1) Note: For simple templates, the pre-denaturation time can be controlled at 30 s-1 min, for complex templates such as bacterial fluids, the pre-denaturation time can be increased to 2 min.
Optimization of parameter settings
1. Template DNA amount setting:
Excessive amounts of template may result in non-specific amplification or smear. The recommended amount of template DNA in a 50 μl PCR reaction system is as follows:
-Human genomic DNA 5 ng-500 ng
-Escherichia coli genomic DNA 50 pg-100 ng
-plasmid DNA 10 pg-1 ng 1. 30-35 number of cycles
2 . Primer concentration setting: The primer concentration can be set between 0.1 μM and 1.0 μM. A low primer concentration may result in low amplification products. Too high a primer concentration will inhibit specific amplification and may result in non-specific amplification.
3. Annealing temperature setting: In general, the annealing temperature is 5℃ lower than the melting temperature of amplification primer Tm, so the annealing temperature can be lowered appropriately when the desired amplification efficiency cannot be obtained; the annealing temperature can be raised appropriately when non-specific reaction occurs. For complex templates, it is necessary to adjust the annealing temperature to achieve efficient amplification.
4. Extension time setting: The extension time should be set according to the size of the amplified fragments. The following extension times are recommended: simple templates such as plasmids: 5-15 s/kb; regular genomes, cDNA templates: 10-15 s/kb; complex templates, crude templates: 20-30 s/kb; (the extension time should not be too short and should be at least 5 s/kb, but should not exceed 30 s/kb).
5. Number of cycles: The number of cycles can be set according to the downstream application of the amplified product. If the number of cycles is too low, the amount of amplification will be insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield.
规格
别名
2×Flash PCR MasterMix（染料）
英文名称
2×Flash PCR MasterMix (Dye)
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
产品线
biological_products
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
技术文档和文章
溶液计算器
常见问题
本产品应如何储存？
请于?20 °C条件下保存。避免反复冻融。建议按单次用量分装保存。
本产品如何运输？
本产品采用保温箱配冰袋运输。收货后请立即拆包，并按上述储存条件保存。
如何获取批次专用COA？
每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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