摘要
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。
A simple and sensitive method for the determnation of deoxyribonucleic acid (DNA) and ribonucleici acid (RNA) has been developed, Our experimental resuits showed that RNA could present fluorescence reaction with terbium(Ⅲ)directly, but DNA presents similar fluorescence reaction only after its denaturation, Dcmaturation occurred in hydrochloric acid solution at pH 2.0～2.8. The optimal conditions of the fluorescence reaction of denatured DNA and RNA with terbium(Ⅲ) have been described. The determination range of acid denatured DNA or RNA was 0.5～20μg/ml and the limit of the sensitivity was about 0.02μg/ml. The relative standard deviation (6 replicateswas) was within 5.0％ in the linear range. Furthermore, comparing native calf thymus (CT) DNA, fish sperm DNA, yeast RNA with acid denatured DNAs and RNA showed that acid denaturation of DNA increased their ability to enhance fluorescence reaction with terbium(Ⅲ) by more several ten-fold than native ones, while it of yeast RNA has little influence on its ability toance, which reflected the difference of Secondary structure of DNAs and RNA. This differencne could be used in the determination of one in the presence of another one.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
1992年第9期1083-1087,共5页
Chinese Journal of Analytical Chemistry
关键词
铽离子
酸变性
荧光增强
DNA
RNA
Deoxyribonucleic acid
Ribonucleic acid
Terbium(Ⅲ)
Acid denaturation
Fluorescence enhancement.
