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Application of a Red-Region Fluorescence Dye,Nile Blue, in Nucleic Acids Assay

Abstract

基于dnA或rnA对耐尔蓝荧光的猝灭效应, 建立一种新的dnA 或rnA 的测定方法. 在最优条件下, 测定小牛胸腺dnA、鲑鱼精子dnA、鲱鱼精子dnA 和酵母rnA 的线性范围分别为: 3-0 ~2-0 x103ng/Ml、9-0 ~2-0 x103ng/Ml、5-4 ~5-0 x103ng/Ml 和27 ~10 x103ng/Ml.检测限分别是3-0ng/Ml、9-0ng/Ml、5-4ng/Ml和27ng/Ml, 相对标准差(n = 6) 是2-1 % . 本文还讨论了干扰物质的影响.A novel fluorimetric method was proposed for the rapid determination of DNA and RNA based on their quenching effect on the cationic red-region fluorescent dye,nile blue.Under optimum conditions,the calibration curves for the determination of Calf thymus DNA(CT DNA),Salmon DNA(SM DNA),Herring sperm (HS DNA) and yeast RNA were linear over the range of 3 0-2 0×10 3 ng/mL,9 0-2 0×10 3 ng/mL,5 4-5 0×10 3 ng/mL and 27-10×10 3 ng/mL,respectively.The detection limits were 3 ng/mL for CT DNA,9 0 ng/mL for SM DNA,5 4 ng/mL for HS DNA and 27 ng/mL for RNA,respectively.The relative standard deviation(n=6) was within 2 1% in the middle of linear range.Interference concerning some interesting coexisting substances with the determination of DNA was also examined.国家自然科学基金!(29775021

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