阿的平荧光猝灭法测定 DNA(英文)

Spectrofluorometric determination of DNA with atebrine

在室温条件下 ,阿的平能嵌入 DNA的碱基对之间 ,而阿的平荧光大幅猝灭 .酸性介质下 ,激发波长 348,4 2 8或 4 4 9nm,碱性介质下激发波长为 36 2或 4 2 2 nm,发射波长均为 5 0 2 nm.基于此对核酸进行定量测定 ,方法简单、快速、灵敏 .对于双链或单链小牛胸腺 DNA,线性范围 0～ 4μg/ m L,检测限 0 .0 3μg/ m L,共存物质干扰小 .在不同浓度 ct DNA存在下 。

The fluorescence quenching reaction of atebrine with DNA was studied. Studies involving natural calf thymus DNA and denatured calf thymus DNA, revealed that double-stranded and single-stranded DNA are capable of quenching the fluorescence of atebrine. Based on this, a sensitive determination method of DNA was developed. The determination can be made both under acidic and alkaline conditions. whether in acidic or alkaline solution, the emission wavelength was 502 nm. But the excitation wavelength was at 348 or 428 or 449 nm when under acidic condition, and at 362 or 422 nm under alkaline condition. Under optimal conditions, the calibration graphs were linear between 0 and 4.0 μg/mL for double-stranded or single-stranded calf thymus DNA. The corresponding detection limit was 0.03 μg/mL. The effect of temperature on the fluorescence intensity of atebrine in the presence of various concentrations of ct DNA showed that the quenching process followed the static mechanism.