Studies On Interaction Of Orange G And Eosin B With DNA And Its Applications By RLS Technique

Based on a number of documents, combined with the laboratory conditions, this thesis select the new reagents that are not reported as DNA resonance light scattering probe. Observed the resonance light scattering spectra of orange G and DNA,eosin B and DNA interactions, and based on a linear relationship with the concentration of DNA and resonance light scattering signal changes value, set up two kinds of sensitive method for determination of trace DNA. These methods have the advantages of simple, rapid, and low detective limits. Used for the determination of actual samples, the result is satisfactory. Discussed the mechanism of action of two kinds of biological stain and DNA combined with absorption spectrometry. The thesis is composed of three parts:1. An overview of the research background, research progress in nucleic acid quantitative analysis, application of resonance light scattering technique in the analysis of nucleic acid and the characteristics and significance of the thesis.2. Study on the resonance light scattering spectra (RLS) of the interaction of biological dye orange G (OG) and DNA. Found in the neutral environment, the addition of DNA can enhance OG RLS intensity, the resonance light scattering characteristic peak at358nm, and in certain conditions, the degree of enhancement linearly with the concentration of DNA. We observed reaction conditions and the influences of coexisting substances. The linear range is0.053～1.20μg·mL-1, the detection limit is0.0159μg·mL-1. The linear regression equation for the method is△I=473.23c (c:μg·mL-1), the related coefficient is R2=0.9984.The method has been applied to the determination of DNA in soybean leaves and astragalus membranaceus leaves, the recovery rate was97.7%-103.0%.3. Studied the influence of DNA on resonance light scattering (RLS) of eosin B. The experiment found that, under acidic condition (pH=3.4), eosin B molecules can be inserted into the double helix structure of DNA, thus increase the RLS intensity, the resonance light scattering characteristic peak at602nm, based the linear relationship between RLS enhancement degree and the addition amount of DNA, established a new method for determination of DNA. We observed reaction conditions and the influences of coexisting substances. The linear range is0.039～14μg·mL-1, the detection limit is0.0118μg·mL-1. The linear regression equation for the method is△I=64.334c(c:μg· mL-1), the related coefficient is R2=0.9979.The method has been applied to the determination of DNA in actual samples, the recovery rate was98.5%～102.2%.