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
本品为由GoldStar Best 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×，具有操作简便快速、灵敏度高、特异性强、稳定性好的优点，可最大限度地减少人为误差和污染。该聚合酶具有5′-3′DNA聚合酶活性，5′-3′外切酶活性和3′-5′外切酶活性，在普通PCR条件下，与GoldStar DNA Polymerase相比，具有扩增效率高、错配率低的优良性能。化学修饰后的酶在常温下没有聚合酶活性，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，该条件可以整合入已有的PCR热循环程序。优化的缓冲体系使酶的作用发挥最大功效，实现对目的片段的高保真、高特异性、高扩增效率、高灵敏度扩增。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。扩增得到的大部分PCR产物3′端附有一个“A”碱基，因此可直接用于T/A克隆。适用于常规的PCR反应和对高保真性有要求的基因克隆等实验。
注意：2×GoldStar Best MasterMix含有GoldStar Best DNAPolymerase, 3.4 mM MgCl2 和 400 µM each dNTP。
Notice: 2×GoldStar Best MasterMix contains GoldStar Best DNA Polymerase, 3.4 mM MgCl2 and 400 μM each dNTP. |
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；2-8℃存放三个月，活性无明显改变。
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
|
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
|
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 Best 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。
This product is a premixed system composed of GoldStar Best DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. It has the advantages of simple and fast operation, high sensitivity, strong specificity, and good stability, which can minimize human error and pollution. This polymerase has 5 '-3' DNA polymerase activity, 5 '-3' exonuclease activity, and 3 '-5' exonuclease activity. Under normal PCR conditions, compared with GoldStar DNA polymerase, it has excellent performance of high amplification efficiency and low mismatch rate. The chemically modified enzyme does not exhibit polymerase activity at room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature. Enzyme activation requires incubation at 95 ℃ for 10 minutes, which can be integrated into existing PCR thermal cycling programs. The optimized buffer system maximizes the effectiveness of the enzyme, achieving high fidelity, specificity, amplification efficiency, and sensitivity for the target fragment. 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. Most PCR products obtained from amplification have an "A" base attached to the 3 'end, making them suitable for direct use in T/A cloning. Suitable for routine PCR reactions and gene cloning experiments that require high fidelity.
Note: 2 x GoldStar Best MasterMix contains GoldStar Best DNAPolymerase, 3.4 mM MgCl2, and 400 µ M each dNTP.
Notice: 2×GoldStar Best MasterMix contains GoldStar Best DNA Polymerase, 3.4 mM MgCl2 and 400 μM each dNTP. |
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; Storage at 2-8 ℃ for three months showed no significant change 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 template, primer structure, and target fragment size.
1. PCR reaction system
|
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
|
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 Best 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
别名
金牌热启动酶预混液
英文名称
2xGoldStar Best 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
biological_products
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):
输入批号以搜索分析图谱:
溶液计算器
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。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）