基本描述

别名

GoldStar 探针混合物

规格或纯度

5 ml

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

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（CW0932）： Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96等。

需要Low ROX校正的仪器（CW2625）： 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（CW2626）： ABI Prism7000/7300/7700/7900，Eppendorf，ABI Step One/Step One Plus等。

G665832	Component	5 mL	Storage
G665832A	2×GoldStar Probe Mixture	5×1 mL	-20℃. Avoid freeze/thaw cycle.
G665832B	ddH2O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

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 μM¹⁾
Reverse Primer，10 µM	1 μl	0.2 μM¹⁾
Probe，10 µM	1 μl	0.2 μM²⁾
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：

步骤	温度	时间	/
预变性	95℃	10 min¹⁾	/
变性	95℃	15 s	35-40 个循环
退火/延伸 ²⁾	60℃	1 min	35-40 个循环

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。

GoldStar Probe Mixture is a premixed system specifically designed for real-time fluorescence quantitative PCR using probe methods (TaqMan, Molecular Beacon, etc.), with a concentration of 2 x, containing GoldStar Taq DNA Polymerase, PCR Buffer, dNTPs, and Mg2+. The operation is simple and convenient. Mainly used for detecting genomic DNA target sequences and RNA reverse transcription cDNA target sequences, such as gene expression analysis, copy number analysis, SNP genotype analysis, etc., suitable for fluorescence quantification using different types of probe methods. The GoldStar Taq DNA Polymerase contained in this product is a chemically modified, novel and highly efficient hot start enzyme. It has no polymerase activity at room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimerization at room temperature. The enzyme activation requires incubation 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 fluorescence signal and higher sensitivity, which can detect single copy templates. By using this product, a wider linear range can be obtained, resulting in more accurate quantification of the target gene. Suitable for all fluorescence quantitative PCR instruments that do not require ROX as a calibration dye.
ROX dye is used to correct the fluorescence signal error between wells in quantitative PCR instruments, and is generally used in Real Time PCR amplification instruments from companies such as ABI and Stratagene. The excitation optical systems of different instruments vary, so the concentration of ROX dye must be matched with the corresponding fluorescence quantitative PCR instrument.

Instruments that do not require ROX calibration (CW0932): Roche LightCycle 480, Roche LightCyler 96, Bio rad iCyler iQ, iQ5, CFX96, etc.
Instrument requiring Low ROX calibration (CW2625): 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, etc.
Instruments that require High ROX calibration (CW2626): ABI Prism7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, etc.

G665832	Component	5 mL	Storage
G665832A	2×GoldStar Probe Mixture	5×1 mL	-20℃. Avoid freeze/thaw cycle.
G665832B	ddH2O	5×1 mL	-20℃. Avoid freeze/thaw cycle.

Notes:
1. Before use, please gently mix upside down to avoid foaming, and use after briefly centrifugation.
2. Avoid repeated freezing and thawing of this product, as repeated freezing and thawing may cause a decrease in product performance. This product can be stored for a long time at -20 ℃, away from light. If frequent use is required in the short term, it can be stored at 2-8 ℃.
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 μl Reaction system	Final concentration
2×GoldStar Probe Mixture	25 μl	1 ×
Forward Primer，10 µM	1 μl	0.2 μM¹⁾
Reverse Primer，10 µM	1 μl	0.2 μM¹⁾
Probe，10 µM	1 μl	0.2 μM²⁾
Template DNA	2 μl³⁾	/
50×Low ROX or High ROX（optional）⁴⁾	1 μl	1 ×
ddH2O	up to 50 μl	/

Attention:
1) Typically, the primer concentration is 0.2 μ M can achieve good results, ranging from 0.1 to 1.0 μ M serves 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. Please refer to the instrument manual or the specific usage requirements of each fluorescent probe for concentration adjustment during actual use.
3) The amount of DNA template is usually based on 10-100 ng genomic DNA or 1-10 ng cDNA as a reference. Due to the different copy numbers of target genes contained in templates of different species, gradient dilution can be applied to the template to determine the optimal template usage.
4) The excitation optical systems of different instruments vary, and depending on the instrument used for fluorescence quantification, 50 x Low ROX or 50 x High ROX can be added.

2. PCR reaction program
Attention! The pre denaturation reaction of this product must be completed at 95 ℃ for 10 minutes!
Two step PCR

Step	Temperature	Time	/
Pre denaturation	95℃	10 min¹⁾	/
Denaturation	95℃	15 s	35-40 cycles
Annealing/Extension ²⁾	60℃	1 min	35-40 cycles

Attention:
1) The hot start enzyme used in this product must be activated under pre denaturation conditions of 95 ℃ and 10 minutes.
2) It is recommended to use a two-step PCR reaction program. If good experimental results cannot be obtained due to the use of primers with lower Tm values, a three-step PCR amplification can be attempted. The annealing temperature should be set within the range of 56 ℃ -64 ℃ as a 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

g665832_goldstar probe mixture.pdf

质检证书（CoA,COO,BSE/TSE 和分析图谱)

C of A & Other Certificates(BSE/TSE, COO):

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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

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