基本描述

别名

PAGE专用PCR mix

规格或纯度

for PAGE

英文名称

2xEs Taq MasterMix

储存温度

-20°C储存,避免反复冻融

运输条件

超低温冰袋运输

产品介绍

产品内容：

E666027	Component	5 mL	Storage
E666027A	2×Es Taq MasterMix (for PAGE)	5×1 mL	-20°C. Avoid freeze/thaw cycle.
E666027B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

Notes: 2×Es Taq MasterMix contain Es Taq DNA Polymerase, 3 mM MgCl₂ and 400 µM each dNTP.

产品简介：

本品是由EsTaq 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× 。EsTaq DNA Polymerase具有扩增效率高、错配率低的优良性能。独创的MasterMix配方使整个反应体系非常稳定，超过98%的PCR扩增能一次成功，同时复杂模板也能得到有效扩增，并可最大限度地减少人为误差和污染。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。主要适用于常规PCR反应和对高保真性有要求的基因克隆等实验， 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

试剂	50 μl反应体系	终浓度
2×Es Taq MasterMix（for Dye）	25 μl	1×
Forward Primer，10 µM	2 μl	0.4 μM
Reverse Primer，10 µM	2 μl	0.4 μM
Template DNA	<0.5 μg	<0.5 μg/50 μl
ddH2O	up to 50 μl

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

步骤	温度	时间	/
预变性	94℃	2 min	/
变性	94℃	30 s	25-35 个循环
退火	55-65℃	30 s	25-35 个循环
延伸	72℃	30 s	25-35 个循环
终延伸	72℃	2 min	/

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， Es 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。

Product content：

E666027	Component	5 mL	Storage
E666027A	2×Es Taq MasterMix (for PAGE)	5×1 mL	-20°C. Avoid freeze/thaw cycle.
E666027B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

Notes: 2×Es Taq MasterMix contain Es Taq DNA Polymerase, 3 mM MgCl₂ and 400 µM each dNTP.

Product Introduction:
This product is a premixed system composed of EsTaq DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. EsTaq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate. The unique MasterMix formula makes the entire reaction system very stable, with over 98% of PCR amplification successful at once. At the same time, complex templates can also be effectively amplified, and human error and contamination can be minimized to the greatest extent. 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. Mainly suitable for conventional PCR reactions and gene cloning experiments that require high fidelity, 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

reagent	50 μlReaction system	final concentration
2×Es Taq MasterMix（for Dye）	25 μl	1×
Forward Primer，10 µM	2 μl	0.4 μM
Reverse Primer，10 µM	2 μl	0.4 μM
Template DNA	<0.5 μg	<0.5 μg/50 μl
ddH2O	up to 50 μl

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

step	temperature	time	/
Pre denaturation	94℃	2 min	/
denaturation	94℃	30 s	25-35 cycle
anneal	55-65℃	30 s	25-35 cycle
extend	72℃	30 s	25-35 cycle
Final extension	72℃	2 min	/

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, and the amplification efficiency of Es Taq DNA Polymerase 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.

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

e666027_2xes taq mastermix.pdf

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