基本描述

别名

金牌探针法qPCR mix | GoldStar 探针混合物

规格或纯度

High ROX

英文名称

GoldStar Probe Mixture

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

G665762	Component	5 mL	Storage
G665762A	2×GoldStar Probe Mixture	5×1 mL	-20℃. Avoid freeze/thaw cycle.
G665762B	50×High ROX	200 μL	-20℃. Avoid freeze/thaw cycle.
G665762C	ddH2O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

产品简介

GoldStar Probe Mixture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cDNA靶序列检测，如基因表达分析，拷贝数分析，SNP基因型分析等，适用于不同类型探针法荧光定量。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，有效避免了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。适用于无需ROX作为校正染料的所有荧光定量PCR仪。

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(G665762)：

Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96等。

需要Low ROX校正的仪器(G665762)：

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(G665762)：

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

试剂	50 μl反应体系	终浓度
2×GoldStar Probe Mixture	25 μl	1×
Forward Primer，10 µM	1 μl	0.2 μM ¹⁾
Reverse Primer，10 µM	1 μl	0.2 μM ¹⁾
Probe，10 µM	1 μl	0.2 μM ²⁾
Template DNA	2 μl³⁾
50×Low ROX or High ROX（optional）⁴⁾	1 μl	1 ×
ddH₂O	up to 50 μl

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，退火温度请以 56℃-64℃的范围作为设定参考。

Product content

G665762	Component	5 mL	Storage
G665762A	2×GoldStar Probe Mixture	5×1 mL	-20℃. Avoid freeze/thaw cycle.
G665762B	50×High ROX	200 μL	-20℃. Avoid freeze/thaw cycle.
G665762C	ddH2O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

Product Introduction

GoldStar Probe Mixture is a premixed system for 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 and Mg2+, which is easy and convenient to operate. It is mainly used for genomic DNA target sequence and RNA reverse transcription post-cDNA target sequence detection, such as gene expression analysis, copy number analysis, SNP genotype analysis, etc. It is suitable for fluorescence quantification by different types of probe methods. GoldStar Taq DNA Polymerase is a chemically modified, new and highly efficient hot-start enzyme, which has no polymerase activity at room temperature and effectively avoids non-specific amplification caused by non-specific binding of primers and templates or primer dimerization at room temperature, and the activation of the enzyme must be incubated at 95℃ 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 target genes can be obtained with this product. It is suitable for all fluorescent quantitative PCR instruments that do not require ROX as a calibration dye.

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 other companies. 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(G665762):

Roche LightCycler 480, Roche LightCyler 96, Bio-rad iCyler iQ, iQ5, CFX96 and others.

Instruments requiring Low ROX calibration(G665762):

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(G665762):

ABI Prism 7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, and others.

matters needing attention

1. Before use, please mix it gently by turning it up and down, avoid foaming as much as possible, and use it after centrifugation for a short time.

2. Avoid repeated freezing and thawing of this product, repeated freezing and thawing may degrade the product performance. This product can be stored for a long time at -20℃, protected from light. If frequent use is required in the short term, 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

Reagents	50 μl Reaction system	Final concentration
2×GoldStar Probe Mixture	25 μl	1×
Forward Primer，10 µM	1 μl	0.2 μM ¹⁾
Reverse Primer，10 µM	1 μl	0.2 μM ¹⁾
Probe，10 µM	1 μl	0.2 μM ²⁾
Template DNA	2 μl³⁾
50×Low ROX or High ROX（optional）⁴⁾	1 μl	1 ×
ddH₂O	up to 50 μl

Note: 1) Usually, the primer concentration of 0.2μM can get better results, and 0.1-1.0μM can be used as a reference for setting the range. 2) The concentration of the probe used is related to the fluorescent quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the instruction manual of the instrument or the specific requirements of the use of each fluorescent probe for the adjustment of the concentration during the 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 shall be activated by the enzyme under the condition of pre-denaturation 95℃ and 10min.

(2) It is recommended to use two-step PCR reaction program, if you do not get good experimental results due to the use of primers with lower Tm values, etc., you can try to carry out three-step PCR amplification, and the annealing temperature, please use the range of 56 ℃ - 64 ℃ as a setting reference.

安全和危险性（GHS）

SDS英文版

技术规格说明书

技术规格说明书

g665762_goldstar probe mixture (high rox).pdf

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。

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体积

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分子量

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稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

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体积 1 (V1)

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浓度 2 (C2)

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复溶计算器

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体积（添加到小瓶中）

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质量（小瓶）

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所需的复溶浓度

GoldStar Probe Mixture

库存信息

基本描述

安全和危险性（GHS）

技术规格说明书

质检证书（CoA,COO,BSE/TSE 和分析图谱)

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器