基本描述

规格或纯度

用于聚丙烯酰胺凝胶电泳

英文名称

2xTaq 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

T665852	Component	5 mL	Storage
T665852A	2×Taq MasterMix (for PAGE)	5×1 mL	-20℃. Avoid freeze/thaw cycle.
T665852B	ddH₂O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

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

试剂	50 μl反应浓度	终浓度
2×Taq MasterMix（for PAGE）	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，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。

T665852	Component	5 mL	Storage
T665852A	2×Taq MasterMix (for PAGE)	5×1 mL	-20℃. Avoid freeze/thaw cycle.
T665852B	ddH₂O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

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

reagent	50 μl Reaction system	final concentration
2×Taq MasterMix（for PAGE）	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

steps	temperature	time	/
Pre denaturation	94℃	2 min	/
denaturation	94℃	30 s	25-35 cycle
anneal	55-65℃	30 s	25-35 cycle
extension	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. 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.

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

t665852_2xtaq mastermix (for page).pdf

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

=

浓度

x

体积

x

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

x

体积 1 (V1)

=

浓度 2 (C2)

x

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

=

质量（小瓶）

÷

所需的复溶浓度