Gold Multiplex PCR Mix

货号: G666077
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规格
库存
价格
数量
5ml
G666077-5 ml
期货 Stock Image
¥897.90
5ml
G666077-5ml
期货 Stock Image
¥897.90
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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。

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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。

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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。

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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

产品内容:

G666077Component5 mLStorage
G666077A2×Gold Multiplex PCR Mix5×1 mL-20°C. Avoid freeze/thaw cycle.
G666077BddH₂O5×1 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:

G666077Component5 mLStorage
G666077A2×Gold Multiplex PCR Mix5×1 mL-20°C. Avoid freeze/thaw cycle.
G666077BddH₂O5×1 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.

规格

别名
金牌多重 PCR 混合物
英文名称
Gold Multiplex PCR Mix
储存与运输
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输

技术文档

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常见问题

本产品应如何储存?
请于?20 °C条件下保存。避免反复冻融。建议按单次用量分装保存。
本产品如何运输?
本产品采用保温箱配冰袋运输。收货后请立即拆包,并按上述储存条件保存。
如何获取批次专用COA?
每个订单都会随附分析证书(COA),您也可以从账户订单历史中下载。如需订购前的COA,请联系您的客户代表,或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
试剂盒包含哪些组分?
完整组分列表见PDP和试剂盒说明书。个别组分的替换品可单独订购——请联系您的客户代表。