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发明公开
CN105803045A 一种荧光等温核酸扩增方法 无效 - 驳回

一种荧光等温核酸扩增方法
摘要：
本发明公开了一种荧光等温核酸扩增方法，该扩增方法包含以下成分：Tris缓冲液、醋酸钾、醋酸镁、二硫苏糖醇、聚乙二醇、ATP、dNTPs、磷酸肌酸、单链结合蛋白、重组酶、UvsY蛋白、DNA聚合酶、核酸外切酶、荧光探针和引物组。应用该方法，可对DNA进行实时荧光监测。在此基础上，加入RNA逆转录酶，可实现对RNA的扩增。本发明公开的等温核酸扩增方法步骤如下：(1)提取DNA或RNA；(2)将DNA或RNA加入到上述核酸扩增体系；(3)混合均匀后放入到可荧光检测仪器中，35-45℃下反应，5-30分钟内即可观察到荧光信号。本方法操作简单，反应快速，对操作人员要求低，具有更广泛的应用范围。
摘要（英）：

The invention discloses a fluorescent isothermal nucleic acid amplification method. The amplification method comprises the following ingredients: a Tris buffer, potassium acetate, magnesium acetate, dithiothreitol, polyethylene glycol, ATP, dNTPs, creatine phosphate, a single-chain binding protein, a recombinant enzyme, a UvsY protein, a DNA polymerase, an exonuclease, a fluorescent probe and a primer group. The method can be applied to carry out real-time fluorescence monitoring on DNA; and based on the method, addition of an RNA retrovirus cane realize RNA amplification. The isothermal nucleic acid amplification method comprises the steps as follows: (1) extracting DNA or RNA; (2) adding the DNA or RNA to a nucleic acid amplification system; and (3) mixing evenly, adding the mixture into a fluorescence detection instrument, reacting at 35-45 DEG C for 5 to 30 minutes to observe a fluorescence signal. The method has the advanategs of simple operation, fast response, low requirements for operators, and broader range of applications.

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