共振光散射法测定生物样本脱氧核糖核酸的含量

Resonance Light Scattering Method for Determining the Content of Biological Samples Deoxyribonucleic Acid

目的：建立溴化乙锭（EB）与脱氧核糖核酸（DNA）的共振光散射光谱法测定生物样本DNA的含量。方法：在含一定量EB的PBS磷酸缓冲溶液（p H 3.5）中,依次加入不同浓度的DNA,检测该体系的共振光散射强度。结果：在波长为363.0 nm处,存在一共振光散射增强峰,其增强强度（IRLS=I-I0）与DNA浓度呈明显的线性关系。线性方程为ΔIRLS=2.8711CDNA＋5.435,R=0.9955,方法的线性范围为0.025~50.8μg/m L,检出限为0.5 ng/m L。结论：本实验建立的EB-DNA共振光散射法是一种灵敏高、适用于生物样本中DNA含量的简便方法。

Objective： To establish the resonance light scattering spectroscopy method of ethidium bromide（EB） and deoxyribonucleic acid（DNA） for measureing DNA content in biological samples. Methods： Using resonance light scattering（RLS） method, the RLS signals of the systems containing amount EB and different concentrations of DNA were detected in p H 3.5 PBS buffer solution. Results： At a wavelength of 363.0 nm, there is an enhanced resonance light scattering peak which was remarkably increases after the addition of DNA. The results showed a linear relationship between the enhanced RLS intensity of the EB-DNA system and DNA concentration. The linear regression equation is represented as follows： ΔIRLS=2.8711CDNA＋5.435,with regression coefficient R=0.9955. The detection limit was 0.5 ng m L-1. Conclusion： The method established of EB-DNA resonance light scattering is a highly sensitive, simple and suitable for determining DNA content in biological samples.