铝试剂荧光光谱法测定核酸

Fluorescence determination of DNA using aurintricarboxylic acid

对铝试剂在不同pH值下的荧光光谱特征进行了研究,发现pH4.04的Britton Robinson缓冲介质中,DNA使铝试剂的荧光光谱发生紫移,且荧光强度与DNA的量呈线性关系。确定了最佳反应条件,DNA、ctDNA、yRNA、fsDNA的工作曲线分别为ΔF=18.4ρ+4.9、ΔF=11.7ρ+5.6、ΔF=9.5ρ+22.8、ΔF=7.9ρ+23.5(μg mL),其线性范围宽,灵敏度较高,并可用于实际样品的测定。

The enhancement of the fluorescence due to the interaction of aurintricarboxylic acid (ATA) with DNA at pH 4.04 Britton-Robinson was investigated. It is considered that the interaction between ATA and DNA generates complex, at the same time that the emission spectrum of ATA-BSA show a red shift. The increase of fluorescence intensity of the complex is linear with amount of DNA. The determination conditions were optimized and the linear equations were determined respectively to deoxyribonucleic acid, yeast ribonucleic acid, fish sperm deoxyribonucleic acid and calf thymus deoxyribonucleic acid, and they were ΔF=18.4ρ+4.9, ΔF=11.7ρ+5.6, ΔF=9.5ρ+22.8, ΔF=7.9ρ+23.5 (μg/mL), respectively. A sensitive and simple method of fluorescence was established for the determination of DNA. This method has been applied to determination of DNA in samples with satisfactory results.