Studies On Determination Of DNA By Resonance Light-scattering (RLS) Technique And Their Applications

Based on a lot of documents and condition of the laboratory, the Resonance light-scattering (RLS) spectra characteristics of interaction between organic small molecule with deoxyribonucleic acid (DNA) is studied, Base on the variation of the intensity of RLS is ratio to the concentration of the DNA establishes four new determination methods of DNA by means of Resonance light-scattering (RLS) technique in this paper.This paper has four chapters.In the first chapter,the RLS spectra characteristics of interaction between Acridine yellow (AY) with DNA was studied by Resonance RLS method in the worth of pH 5.5 medium. The effects of interference substances and optimization of procedure were investigated and a new method for determination of the DNA was established .It based on RLS method, The linear relationships were studied between the decreased intensity of RLS at 436nm and concentration of deoxyribonucleic acid in the AY solution of two concentrations, good correlation coefficient 0.9996 and 0.9906 were obtained for the concentration of AY 2.0×10-5mol/L and 1.0×10-5mol/L respectively,the detection limits are 10.69μg/L for the concentration of AY 2.0×10-5mol/L and 7.43μg/L for the concentration of AY 1.0×10-5mol/L. The approach has good operation, high sensitivity and accuracy The widest linear range of the DNA is 0-1.25mg/L. The method has been applied to determination of fish sperm DNA with satisfactory results.In the second chapter, A new method for the determination of the DNA was established based on RLS technique. Characteristics of interaction between Nitro-tetrazolium chloride blue(NCB) and DNAwere studied in the value of pH 1.8 medium by RLS technique. The effects of interference substances and reaction condition on system were investigated and the optimal condition was presented.The linear relationships were studied between the enhanced intensity of RLS at 436nm and concentration of deoxyribonucleic acid in the NCB solution of two concentrations, good correlation coefficient 0.9958 and 0.9965 were obtained for the concentration of NCB 4.0×10-5mol/L and 2.0×10-5mol/Lrespectively, the detection limits are 30.04μg/L for the concentration of NCB 4.0×10-5mol/L and 67.57μg/L for the concentration of NCB 2.0×10-5mol/L. The approach has good operation, high sensitivity and accuracy and wide determination range.It has been applied to determination of fish sperm DNA with satisfactory results.In the third chapter, the RLS spectra Characteristics of interaction between Alkali blue 6B with DNA was studied by Resonance RLS method in the worth of pH 2.90 medium. The effects of interference substances and optimization of procedure were investigated and a new method based on RLS was established .The method can determine the DNA at nanogram level. The linear range of the DNA is 10～100.4μg/L with a correlation coefficient of 0.9913. The detection limit is 4.2μg/L. It has been applied to determination of fish sperm DNA with satisfactory results. In the fourth chapter, A new method for determination of nucleic acid has been established. Which base on nucleic acids and CTMAB can enhance the Resonance Light-Scattering (RLS) intensity in existing Xylene Cyanal FF, in the worth of pH 9.9's Buffer-solution of Tris-HCl. The relationships between the enhancement of RLS intensity and the concentration of three nucleic acids were studied, Under the optimal conditions. The linear rang are 0.05～3.0mg/L,0.05～6.0mg/L,0.5～2.0mg/L for the fsDNA, ctDNA, and yRNA respectively. The detection limits are 24.1μg/L. 18.4μg/L. 57.0μg/L respectively. The method has been applied to determination of fsDNA in mixed samples with satisfactory results.