DNA荧光探针—荧光素-中性红体系的研究

Fluorescein-Neutral Red Pair as a New Donor-acceptor Set for Fluorescence Energy Transfer Study of DNA

  • 摘要: 基于DNA对荧光素(FL)-中性红(NR)分子间荧光能量转移的抑制作用,以荧光素-中性红为荧光探针,考察该探针与DNA的结合反应,建立了准确测定DNA的新方法,在pH=6.5条件下,hsDNA、ctDNA和smDNA的浓度与荧光素-中性红体系的荧光比值变化量Δ(Fd/Fa)成线性关系,响应线性范围分别为0.25~6.25μg/mL、0.10~5.00μg/mL、0.10~4.00μg/mL,检测限(3σ/K)分别为0.021μg/mL、0.025μg/mL和0.023μg/mL;分析测定了DNA合成样品,回收率91.3%~101.4%,相对标准偏差小于4.2%.

     

    Abstract: After the interaction of fluorescein and neutral red with DNA, the fluorescence transfer system developed for nucleic acids had been studied by using fluorimetric, ultra-violet methods. Experiments revealed that nucleic acids could enhance remarkably the fluorescence intensity of Fluorescein with fluorescence quenching of neutral red in the presence of DNA. Under optimum conditions, the calibration graphs are linear over the range of 0.25~6.25 μg/mL for herring sperm DNA, 0.10~4.00 μg/mL for salmon DNA, 0.10~5.00 μg/mL for calf thymus DNA with the detection limit (3σ/K) being 0.021 μg/mL, 0.023 μg/mL and 0.025 μg/mL respectively. In the determination of nucleic acids in synthetic samples, the value of RSD for several replicates is less than 4.2% and the recoveries range between 91.3% and 101.4%.

     

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