用于聚丙烯酰胺凝胶电泳 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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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。
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注意：2×Taq MasterMix含有Taq DNA Polymerase, 3 mM MgCl2 和400 µM each dNTP。
产品简介：
2×Taq MasterMix是由Taq DNA Polymerase、Mg2+、dNTPs、PCR稳定剂和增强 剂组成的预混体系。预配好的PCR混合液使操作更加简单快捷，并可最大限度地减少 人为误差和污染。独创的MasterMix配方使扩增产物得率高，重复性强，稳定性好。本 品不含染料，PCR程序结束后可根据需要加入适量上样缓冲液后进行电泳操作。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。主要适用于PCR法扩 增DNA、DNA序列测定等实验，PCR扩增产物专用于聚丙烯酰胺凝电泳检测。
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。
1. PCR反应体系
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2. PCR反应条件
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注意：
1）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，适当降 低退火温度；发生非特异性反应时，提高退火温度，由此优化反应条件。
2）延伸时间应根据所扩增片段大小设定，本产品Taq DNA Polymerase的扩增效率为2 kb/min。
3）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。
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Attention:2x Taq MasterMix contains Taq DNA polymerase, 3 mM MgCl2, and 400 µ M per dNTP
Product Introduction:
2x Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, PCR stabilizers, and enhancers. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent possible. The original MasterMix formula results in high yield, strong repeatability, and good stability of amplified products. This product does not contain dyes. After the PCR program is completed, an appropriate amount of sample buffer can be added as needed for electrophoresis operation. The amplified PCR product has an "A" base attached to its 3 'end, making it suitable for direct use in T/A cloning. Mainly suitable for PCR amplification of DNA, DNA sequencing and other experiments, PCR amplification products are specifically used for polyacrylamide gel electrophoresis detection.
Quality control:
After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.
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. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.
1. PCR reaction system
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Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration μ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.
2. PCR reaction conditions
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Attention:
1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.
2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Taq DNA Polymerase in this product is 2 kb/min.
3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible.
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