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本文提出了一种简单灵敏的测定DNA和RNA的方法。实验结果表明RNA可直接与铽离子发生荧光反应,但DNA只有在发生变性之后才能发生同样的反应。酸变性的最佳pH范围在2.2～2.8之间,通过实验确定了荧光反应的最佳条件。在最佳条件下测定DNA和RNA的线性范围在0.5～20.0μg/ml,标准偏差在5％以內。小牛胸腺DNA和鱼精子DNA在酸变性时其对铽离子荧光增强效应提高数十倍,而RNA变性前后对铽离子荧光增强效应变化不大,这本身反映了DNA与RNA二级结构的不同。利用DNA和RNA在酸变性时对Tb(Ⅱ)荧光增强变化的不同可在一定量的DNA存在下测定RNA或在一定量的RNA存在下利用差减法测DNA。
This article presents a simple and sensitive method for the determination of DNA and RNA. The experimental results show that RNA can directly react with terbium ions, but the same reaction occurs only after DNA denaturation. The optimum pH range of acid denaturation is between 2.2 and 2.8. The optimum conditions of fluorescence reaction were determined experimentally. Under the optimum conditions, the linear range of DNA and RNA was 0.5 ~ 20.0μg / ml, and the standard deviation was within 5%. Calf thymus DNA and fish sperm DNA enhances the fluorescence enhancement effect of terbium ions by several tens of times when acid denatured, while the fluorescence enhancement effect of terbium ions does not change much before and after RNA denaturation, which itself reflects the difference of DNA and RNA secondary structure . The use of DNA and RNA to alter Tb (II) fluorescence during acid denaturation can be used to determine RNA in the presence of a quantity of DNA or to subtract to subtract the DNA in the presence of a defined amount of RNA.