产品内容：

产品简介：

Gold  Multiplex  PCR  Mix，是由GoldStar  DNA  Polymerase、Mg2+ 、dNTPs以及 PCR稳定剂和增强剂等组成的预混体系。使用本产品无需进行繁杂的PCR反应条件的 优化过程，只需进行简单的条件摸索即可轻松进行多重PCR反应。本品中所含的 GoldStar  DNA  Polymerase是经化学修饰的热启动酶，可以有效减少PCR反应初期因 引物错配而产生的非特异扩增。酶的激活须在95°C下孵育10分钟此酶与能提高反应特 异性的PCR增强剂以及独特的缓冲体系相配合，使反应体系中所有的引物都能有效延伸， 无需额外优化。此MasterMix中还包含GC Enhancer，有助于实现“ 困难”模板（比如高 GC含量的模板）的高效扩增。 Gold  Multiplex  PCR  Mix适用于各种类型的多重PCR反 应，比如微卫星分析、基因分型和SNP检测等。

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；2-8℃存放三个月，活 性无明显改变。

使用方法：

以下举例为常规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

注意：引物设计时，尽量减小各引物的Tm间的差值，差值尽量控制在5℃以内。各引物浓度请 以终浓度0.05-0.2 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 非特异性扩增时，可降低引物浓度，由此优化反应体系。

2. PCR反应条件

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，本产品中所包含的GoldStar DNA Polymerase的扩增效 率为1-2 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。

4）本产品在预变性95°C，10 min条件下实现酶的活化。

Product content:

Product Introduction:
Gold Multiplex PCR Mix is a premixed system composed of GoldStar DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers. Using this product does not require a complicated optimization process for PCR reaction conditions, and multiple PCR reactions can be easily carried out with simple condition exploration. The GoldStar DNA Polymerase contained in this product is a chemically modified hot start enzyme that can effectively reduce non-specific amplification caused by primer mismatch in the early stages of PCR reactions. The activation of the enzyme needs to be incubated at 95 ° C for 10 minutes. This enzyme, in combination with PCR enhancers that can enhance reaction specificity and a unique buffer system, allows all primers in the reaction system to be effectively extended without the need for additional optimization. This MasterMix also includes GC Enhancer, which helps achieve efficient amplification of "difficult" templates, such as those with high GC content. Gold Multiplex PCR Mix is suitable for various types of multiplex PCR reactions, such as microsatellite analysis, genotyping, and SNP detection.

quality control：
After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Storage at 2-8 ℃ for three months showed no significant change in activity.

Usage：
The following are examples of conventional PCR reaction systems and reaction conditions. In practical operation, corresponding improvements and optimizations should be made based on different templates, primer structures, and target fragment sizes.

1. PCR reaction system

Attention: When designing primers, try to minimize the difference in Tm between each primer, and control the difference within 5 ℃ as much as possible. Please use the final concentration of 0.05-0.2 for each primer 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 amplification occurs, the primer concentration can be reduced to optimize the reaction system.

2. PCR reaction conditions

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 GoldStar DNA Polymerase contained in this product is 1-2 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.
4) This product achieves enzyme activation under pre denaturation conditions of 95 ° C for 10 minutes.