计算溶液所需的质量、体积或浓度。
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。
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。
质量文档
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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
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产品简介
2×GoldStar Probe Mixture（UNG）是专用于探针法（TaqMan，Molecular Beacon 等）实时荧光定量PCR的预混体系，浓度为2×，包含GoldStar Taq DNA polymerase、PCR Buffer、dNTPs（dTTP全部被dUTP所取代）、UNG酶和Mg2+，操作简单方便。主要用于基因组DNA靶序列和RNA反转录后的cDNA靶序列的检测，如基因表达分析， 拷贝数分析，SNP基因型分析等。本产品中运用了dUTP-UNG防污染系统，在PCR反 应体系配制过程中加入了dUTP，因此就形成了含有dU碱基的扩增产物。而此产物可以 在下次进行PCR反应前，由PCR体系中的UNG酶处理消除。这样就有效的去除了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。本品含有 的GoldStar Taq DNA Polymerase是一种经化学修饰的、全新高效热启动酶，在常温下没有聚合酶活性，有效避免在常温条件下由引物和模板非特异性结合或引物二聚体而产 生的非特异性扩增，酶的激活须在95℃下孵育10分钟。独特的PCR缓冲体系与热启动 酶的组合，显著提高了PCR的扩增效率，荧光信号更强，灵敏度更高，可以检测单拷贝 的模板。使用本产品可以得到更广的线性范围，对目的基因定量更准确。
ROX染料用于校正定量PCR仪孔与孔之间产生的荧光信号误差，一般用于ABI、Stratagene等公司的Real Time PCR 扩增仪。不同仪器的激发光学系统有所不同，因此ROX染料的浓度必须与相应的荧光定量PCR仪相匹配。
不需要ROX校正的仪器 （G670150） ：
Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96等。
需要Low ROX校正的仪器 （G670150） ：
ABI Prism7500/7500 Fast，QuantStudio® 3 System，QuantStudio® 5 System， QuantStudio® 6 Flex System，QuantStudio® 7 Flex System，ViiA 7 system， Stratagene Mx3000/Mx3005P，Corbett Rotor Gene 3000等。
需要High ROX校正的仪器 （G670150） ：
ABI Prism7000/7300/7700/7900，Eppendorf，ABI Step One/Step One Plus等。
注意事项
1．使用前请上下颠倒轻轻混匀，尽量避免起泡，并经短暂离心后使用。
2．避免反复冻融本品，反复冻融可能使产品性能下降。本产品长期保存可置于-20℃， 避光。如果在短期内需要频繁使用，可在2-8℃保存。
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
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注意：1）通常引物浓度以0.2 μM可以得到较好结果，可以在0.1-1.0 μM作为设定范围的参考。
2）使用的探针浓度，与使用的荧光定量PCR仪、探针种类、荧光标记物质种类有关，实际使 用时请参照仪器说明书，或各荧光探针的具体使用要求进行浓度的调节。
3）通常DNA模板的量以10-100 ng基因组DNA或1-10 ng cDNA为参照，因不同物种的模板中含 有的目的基因拷贝数不同，可对模板进行梯度稀释，以确定最佳的模板使用量。
4）不同仪器的激发光学系统有所不同，根据使用荧光定量的仪器选择加入50× Low ROX or
50×High ROX。
2. PCR反应程序
注意！本产品预变性反应必须在95℃ 10分钟下完成！
两步法PCR：
注意：1）本产品所采用的热启动酶须在预变性95℃、10 min条件下实现酶的活化。
2）建议采用两步法PCR反应程序，若因使用Tm值较低的引物等原因，得不到良好的实验结果 时，可尝试进行三步法PCR扩增。
三步法PCR：
Product content
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Product Introduction
2× GoldStar Probe Mixture (UNG) is a premixed system dedicated to real-time fluorescence quantitative PCR by probe method (TaqMan, Molecular Beacon, etc.), with a concentration of 2×, containing GoldStar Taq DNA polymerase, PCR Buffer, dNTPs (dTTP is all replaced by dUTP), UNG enzyme and Mg2+, which is easy and convenient to operate. It is mainly used for the detection of genomic DNA target sequences and cDNA target sequences after RNA reverse transcription, such as gene expression analysis, copy number analysis and SNP genotype analysis. This product utilizes the dUTP-UNG anti-pollution system, which adds dUTP during the preparation of the PCR reaction system, thus forming an amplification product containing dU bases. This product can be eliminated by the UNG enzyme in the PCR system before the next PCR reaction. This effectively removes residual contamination of the PCR product and greatly reduces false positives due to contamination of the amplification product.UNG enzyme can be inactivated at the pre-denaturation step in the PCR cycle, and therefore will not affect the formation of new PCR products containing dU bases. The GoldStar Taq DNA Polymerase contained in this product is a chemically modified, new high-efficiency hot-start enzyme, which has no polymerase activity at room temperature, effectively avoiding non-specific amplification due to non-specific binding of primers and templates or primer dimerization at room temperature, and the activation of the enzyme must be incubated at 95°C for 10 minutes. The unique combination of PCR buffer system and hot-start enzyme significantly improves the amplification efficiency of PCR with stronger fluorescent signal and higher sensitivity to detect single-copy templates. A wider linear range and more accurate quantification of the target gene can be obtained by using this product.
ROX dye is used to correct the fluorescence signal error generated between wells of a quantitative PCR instrument, and is generally used in Real Time PCR amplifiers from ABI, Stratagene, and others. The excitation optics vary from instrument to instrument, so the concentration of ROX dye must be matched to the corresponding fluorescence quantitative PCR instrument.
Instruments that do not require ROX calibration （G670150） :
Roche LightCycler 480, Roche LightCyler 96, Bio-rad iCyler iQ, iQ5, CFX96 and others.
Instruments requiring Low ROX calibration （G670150） :
ABI Prism7500/7500 Fast, QuantStudio®3 System, QuantStudio®5 System, QuantStudio®6 Flex System, QuantStudio®7 Flex System, ViiA 7 system. Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000, and more.
Instruments requiring High ROX calibration （G670150） :
ABI Prism 7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, and others.
matters needing attention
1. Before use, please mix gently by turning up and down, avoid foaming as much as possible, and use after brief centrifugation.
2. Avoid repeated freezing and thawing of this product, repeated freezing and thawing may degrade the product performance. This product can be stored for long term at -20℃, protected from light. If frequent use is required within a short period of time, it can be stored at 2-8℃.
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
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Note: 1) Usually, better results can be obtained with a primer concentration of 0.2 μM, and 0.1-1.0 μM can be used as a reference for setting the range.
(2) The concentration of the probe used is related to the fluorescence quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, please refer to the instrument manual or the specific requirements for the use of each fluorescent probe for the adjustment of the concentration in actual use.
(3) Usually the amount of DNA template is 10-100ng genomic DNA or 1-10ng cDNA as a reference. Since the templates of different species contain different copy numbers of target genes, the templates can be subjected to gradient dilution to determine the optimal amount of template to be used.
(4) The excitation optical system varies from instrument to instrument, choose to add 50×Low ROX or 50×High ROX according to the instrument using fluorescence quantification.
2. PCR reaction program
Caution! The pre-denaturation reaction of this product must be completed at 95°C for 10 minutes!
Two-step PCR:
Note: 1) The hot-start enzyme used in this product must be activated under the condition of pre-denaturation 95℃, 10min.
2) It is recommended to use the two-step PCR reaction program, if you can not get good experimental results due to the use of primers with a lower Tm value, etc., you can try to carry out the three-step PCR amplification.
Three-step PCR:
规格
别名
GoldStar 探针混合物
规格或纯度
UNG+Low ROX
英文名称
GoldStar Probe Mixture
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
小分子
技术文档
高级数据
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
溶液计算器
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。
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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。
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