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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。
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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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
FastSYBR Mixture是专用于染料法(SYBR GreenⅠ)实时荧光定量PCR的预混体系，浓度为2×,包含Fast Taq DNA Polymerase、PCR Buffer、dNTPs、SYBR Green I 荧光染料和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靶序列检测。本品所含荧光染料SYBR Green I可以与所有的双链DNA相结合，使该产品可用于不同靶序列的检测而不需合成特异性标记探针。本品含有的Fast Taq DNA Polymerase能有效减少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，酶的激活仅需在95℃孵育20s。整个PCR反应过程比普通反应可节省约40分钟，大大缩短了PCR的反应时间。独特的PCR缓冲体系与热启动酶的组合， 有效抑制了非特异产物的产生，并显著提高PCR的扩增效率。该产品应用范围广，适用于普通和快速定量PCR程序。
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（F665884） ：
Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96等。
需要Low ROX校正的仪器（F665743） ：
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（F665747） ：
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．本产品中含有荧光染料SYBR Green I，保存本产品或配制PCR反应液时应避免强光 照射。
3．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。
4．本品不能用于探针法荧光定量PCR。
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。 扩增效率不高的情况下，可提高引物的浓度；发生非特异性反应时，可降低引物浓度，由此优 化反应体系。
2）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。
3）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反应条件
注意：1）本产品所采用的酶须在预变性95℃、20s条件下实现酶的活化。在此条件下，大多数 模板可良好的进行解链。对GC含量高、二级结构复杂的模板，可将预变性时间延长至1分钟， 以使起始模板充分解链。若高温处理时间过长，会对酶的活性造成影响。可根据模板情况确定 最佳的预变性时间。
2）建议采用两步法PCR反应程序，退火温度请以60-64℃作为设定范围的参考，发生非特异性 反应时，可提高退火温度。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。
3）融解曲线分析请以所使用的荧光定量PCR仪推荐的程序进行设定，本程序是以ABI 7500荧光
定量PCR仪为参照设定。
Product content
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Product Introduction
FastSYBR Mixture is a premixed system for dye-based (SYBR Green I) real-time fluorescent quantitative PCR, with a concentration of 2×, containing Fast Taq DNA Polymerase, PCR Buffer, dNTPs, SYBR Green I fluorescent dye and Mg2+, which is easy to operate. It is mainly used for the detection of genomic DNA target sequence and cDNA target sequence after RNA reverse transcription. The fluorescent dye SYBR Green I contained in this product can bind to all double-stranded DNAs, enabling the product to be used for the detection of different target sequences without the need to synthesize specific labeling probes. The Fast Taq DNA Polymerase can effectively reduce the non-specific amplification caused by the non-specific binding of primers and templates or primer dimerization at room temperature, and the activation of the enzyme only requires incubation at 95℃ for 20 s. The whole PCR reaction can save about 40 minutes compared with the normal reaction, which greatly shortens the reaction time of PCR. The unique combination of PCR buffer system and hot-start enzyme effectively inhibits the generation of non-specific products and significantly improves the amplification efficiency of PCR. The product has a wide range of applications and is suitable for both normal and rapid quantitative PCR programs.
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 (F665884) :
Roche LightCycler 480, Roche LightCyler 96, Bio-rad iCyler iQ, iQ5, CFX96 and others.
Instruments that require Low ROX calibration (F665743) :
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 that require High ROX calibration (F665747) :
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. This product contains the fluorescent dye SYBR Green I. Avoid strong light when storing this product or preparing PCR reaction solution.
3. 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℃.
4. This product cannot be used for fluorescent quantitative PCR by the probe method.
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. If the amplification efficiency is not high, the concentration of the primer can be increased; when a non-specific reaction occurs, the concentration of the primer can be decreased, thus optimizing the reaction system.
2）Usually the amount of DNA template is 10-100 ng genomic DNA or 1-10 ng cDNA as a reference. Since the number of copies of the target gene contained in the template varies from species to species, the template can be diluted in a gradient to determine the optimal amount of template to be used.
3）The excitation optical system varies from instrument to instrument, so choose whether to add 50×Low ROX or 50×High ROX depending on the instrument used for fluorescence quantification.
2. PCR reaction conditions
Note: 1) The enzyme used in this product must be pre-denatured at 95°C, 20s to achieve enzyme activation. Under this condition, most of the templates can be well unchained. For templates with high GC content and complex secondary structure, the pre-denaturation time can be extended to 1 minute in order to make the starting template fully unchained. If the high temperature treatment time is too long, it will affect the enzyme activity. The optimal pre-denaturation time can be determined according to the template.
(2) It is recommended to use two-step PCR reaction program, the annealing temperature should be 60-64 ℃ as the reference of the setting range, and the annealing temperature can be increased when a non-specific reaction occurs. If you can't get good results due to the use of primers with low Tm value, you can try three-step PCR amplification, and the annealing temperature should be set in the range of 56℃-64℃ as a reference.
(3) For melting curve analysis, please set up the program recommended for the fluorescence quantitative PCR instrument used, and this program is set up using the ABI 7500 fluorescence quantitative PCR instrument as a reference.
规格
别名
FastSYBR 混合物
规格或纯度
High ROX
英文名称
FastSYBR 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):
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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？
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