阿的平荧光猝灭法测定DNA

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348,428或449nm,碱性介质下激发波长为362或422nm,发射波长均为502nm,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/mL,检测限0．03μg/mL,共存物质干扰小,在不同浓度ctDNA存在下,温度对荧光光猝灭的影响结果显示猝灭方式为静态猝灭。

Spectrofluorometric determination of DNA with atebrine

The fluorescence quenching reaction of atebrine with DNA was studied. Studies involving natural calf thymus DNA and denatured calf thymus DNA, reveale d t hat double stranded and single stranded DNA are capable of quenching the fluo rescen ce of atebrine. Based on this, a sensitive determination method of DNA was devel oped . The determination can be made both under acidic and alkaline conditions. Whet her in acidic or alkaline solution, the emission wavelength was 502 nm. But th e e x citation 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 calibra t ion graphs were linear between 0 and 4.0 μg／Ml for double stran ded or single s tranded 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.