基本描述

别名

金牌多重 PCR 混合物

英文名称

Gold Multiplex PCR Mix

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：

G666077	Component	5 mL	Storage
G666077A	2×Gold Multiplex PCR Mix	5×1 mL	-20°C. Avoid freeze/thaw cycle.
G666077B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

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

试剂	50 μl反应体系	终浓度
2×Gold Multiplex PCR Mix	25 μl	1×
Primer Mix，10 µM each	1 μl	0.2 μM
Template DNA	适量	/
ddH2O	up to 50 μl	/

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

步骤	温度	时间	/
预变性	95℃	10 min	/
变性	95℃	30 s	30-40个循环
退火	55-65℃	30 s	30-40个循环
延伸	72℃	1 kb/min	30-40个循环
终延伸	72℃	5 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:

G666077	Component	5 mL	Storage
G666077A	2×Gold Multiplex PCR Mix	5×1 mL	-20°C. Avoid freeze/thaw cycle.
G666077B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

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

reagent	50 μlReaction system	final concentration
2×Gold Multiplex PCR Mix	25 μl	1×
Primer Mix，10 µM each	1 μl	0.2 μM
Template DNA	appropriate amount	/
ddH2O	up to 50 μl	/

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

step	temperature	time	/
Pre denaturation	95℃	10 min	/
denaturation	95℃	30 s	30-40cycle
anneal	55-65℃	30 s	30-40cycle
extension	72℃	1 kb/min	30-40cycle
Final extension	72℃	5 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 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.

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

g666077_gold multiplex pcr mix.pdf

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货号 (SKU)	包装规格	是否现货	价格	数量
G666077-5 ml	5ml	期货
G666077-5ml	5ml	期货