基本描述

规格或纯度

UNG

英文名称

Multiplex PCR 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

M665822	Component	5×1 mL	Storage
M665822A	2×Multiplex PCR MasterMix (UNG）	5×1 mL	-20°C. Avoid freeze/thaw cycle.
M665822B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

产品简介

2×Multiplex PCR MasterMix (UNG）是由GoldStar Taq DNA Polymerase、Mg2+、dNTPs（含dUTP）、UNG酶以及PCR稳定剂等组成的PCR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。

本产品所含的GoldStar Taq 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。独特的缓冲体系，使多重PCR反应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。本产品运用dUTP-UNG 防污染系统，可有效去除了PCR产物的残留污染，大大降低了由于扩增产物污染而导致的假阳性。UNG酶在PCR循环中的预变性步骤即可被灭活，因此不会影响新的含dU 碱基PCR产物的形成

Multiplex PCR MasterMix (UNG）可有效防止PCR产物的残留污染，适用于防污染多重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3天，扩增性能无明显改变。

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×Multiplex PCR MasterMix (UNG)	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0.05-0.2μM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：

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 Taq DNA Polymerase 的扩增效率为1 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

3．结果检测：本产品不含染料，反应结束后，取5 µl反应产物加入适量上样缓冲液后进行电泳检测结果。

Product content

M665822	Component	5×1 mL	Storage
M665822A	2×Multiplex PCR MasterMix (UNG）	5×1 mL	-20°C. Avoid freeze/thaw cycle.
M665822B	ddH₂O	5×1 mL	-20°C. Avoid freeze/thaw cycle.

Product Introduction

2×Multiplex PCR MasterMix (UNG) is a PCR premix system consisting of GoldStar Taq DNA Polymerase, Mg2+, dNTPs (including dUTP), UNG enzyme and PCR stabilizer. This product eliminates the need for complicated optimization of PCR reaction conditions and allows for easy multiplexing of PCR reactions by simple mapping of conditions.

The GoldStar Taq DNA Polymerase contained in this product is a chemically modified hot-start enzyme that effectively reduces non-specific amplification due to primer mismatches at the beginning of the PCR reaction. The unique buffer system allows all primers of the multiplex PCR reaction to be extended efficiently without additional optimization. The MasterMix also includes a GC Enhancer, which helps to achieve efficient amplification of "difficult" templates (e.g., those with high GC content). False positives due to contamination of amplification products are greatly reduced by the use of the dUTP-UNG Anti-Contamination System, which effectively removes residual contamination of PCR products. ung enzyme is inactivated during the pre-denaturation step of the PCR cycle, so that it does not interfere with the formation of new PCR products containing dU bases.

Multiplex PCR MasterMix (UNG) effectively prevents residual contamination of PCR products and is suitable for contamination-proof multiplex PCR reactions such as microsatellite analysis, genotyping and SNP detection.

quality control

No exogenous nuclease activity was examined; no host residual DNA was detected by PCR method; and the amplification performance was not significantly altered by storage at 2-8°C for 3 days.

Usage

The following examples are conventional PCR reaction systems and reaction conditions, which should be improved and optimized according to the template, primer structure and target fragment size in actual operation.

1.PCR reaction system:

Reagents	50 μl Reaction system	Final concentration
2×Multiplex PCR MasterMix (UNG)	25 μl	1×
Primer Mix，10 µM each	1 μl	0.2 μM
Template DNA	appropriate amount
ddH2O	up to 50 μl

Note: When primer design, try to minimize the difference between the Tm of each primer, and the difference should be controlled within 5℃ as much as possible. Please use the final concentration of 0.05-0.2μM as the reference for setting the range of each primer concentration. If the amplification efficiency is not high, the concentration of primers can be increased; when non-specific amplification occurs, the concentration of primers can be decreased, thus optimizing the reaction system.

2.PCR reaction conditions:

Note: 1) In general, the annealing temperature in the experiment is 5°C lower than the melting temperature Tm of the amplification primer, and when the ideal amplification efficiency cannot be obtained, the annealing temperature is appropriately lowered; when a non-specific reaction occurs, the annealing temperature is raised, thus optimizing the reaction conditions.

2）The extension time should be set according to the size of the fragment being amplified. The amplification efficiency of GoldStar Taq DNA Polymerase included in this product is 1kb/min.

3）The number of cycles can be set according to the downstream application of the amplification product. If the number of cycles is too low, the amplification is insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield.

3. Result detection: This product does not contain dyes, after the reaction is finished, take 5µl of the reaction product and add the appropriate amount of sampling buffer for electrophoresis to detect the results.

AI解读

技术规格说明书

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

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