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
2×GoldStar MasterMix是由GoldStar DNA Polymerase、PCR Buffer、Mg2+、 dNTPs及PCR稳定剂和增强剂组成的预混体系，预混好的PCR混合液使操作更加简单 快捷，可最大限度地减少人为误差和污染。该产品所含有的GoldStar DNA Polymerase是 一种经化学修饰的、全新高效Taq DNA Polymerase，在常温下酶的活性被完全封闭 ，使得该酶在低温或常温下没有活性，从而有效避免在常温条件下由引物和模板非特 异性结合或引物二聚体而产生的非特异性扩增，酶的激活须在95℃下孵育10分钟。独 特的缓冲体系使酶的应用更为广泛，使GC含量高、二级结构复杂以及低拷贝的模板，均 能高效扩增。特有的MasterMix配方使整个反应体系稳定性更强。用本品进行PCR扩增 ，PCR产物3'末端附有一个“A”碱基，可直接用于T/A克隆。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。本产品特异性强，PCR扩增后 无需胶回收去除杂带，可直接用于下游克隆或芯片杂交等实验。主要应用于常规的 PCR、RT-PCR、多重PCR和基因芯片检测，特别适用于对特异性要求较高的PCR反应。
Notes: 2×GoldStar Taq MasterMix contains GoldStar DNA Polymerase, 3.4 mM MgCl2 and 400 µM each dNTP. |
质量控制
经检测无外源核酸酶活性；PCR方法检测无宿主残留DNA；能有效扩增人基因组 中的单拷贝基因；2-8°C存放三个月，无明显活性改变。
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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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。
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℃，退火时间一般为30-60 s，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）本产品须在预变性95℃，10 min条件下实现酶的活化。
2 x GoldStar MasterMix is a pre mixed system composed of GoldStar DNA Polymerase, PCR Buffer, Mg2+, dNTPs, PCR stabilizers and enhancers. The pre mixed PCR mixture makes the operation simpler and faster, minimizing human error and contamination. The GoldStar DNA Polymerase contained in this product is a chemically modified, novel and highly efficient Taq DNA Polymerase. The enzyme's activity is completely blocked at room temperature, making it inactive at low or room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature. The activation of the enzyme requires incubation at 95 ℃ for 10 minutes. The unique buffer system makes the application of enzymes more extensive, enabling efficient amplification of high GC content, complex secondary structures, and low copy templates. The unique MasterMix formula enhances the stability of the entire reaction system. Using this product for PCR amplification, the 3 'end of the PCR product contains an "A" base, which can be directly used for T/A cloning. 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. This product has strong specificity and does not require gel recovery to remove impurities after PCR amplification. It can be directly used for downstream cloning or chip hybridization experiments. Mainly used for routine PCR, RT-PCR, multiplex PCR, and gene chip detection, especially suitable for PCR reactions that require high specificity.
Notes: 2×GoldStar Taq MasterMix contains GoldStar DNA Polymerase, 3.4 mM MgCl2 and 400 µM each dNTP. |
Quality control
No exogenous nuclease activity detected; PCR method for detecting host free residual DNA; Can effectively amplify single copy genes in the human genome; Store at 2-8 ° C for three months without significant changes in activity.
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 different template primer structures and target fragment sizes.
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, the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the annealing time is generally 30-60 seconds. If 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 must achieve enzyme activation under pre denaturation conditions of 95 ℃ and 10 minutes.
规格
英文名称
2xGoldStar MasterMix
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
生物产品
名称和识别符
分子类型
小分子
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):
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溶液计算器
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