Studying And Application Of Resonance Light Scattering Technique In Nucleic Acids Analysis

Nucleic acid is the carrier of genetic information and the material basis of geneexpression. It plays an important role in the biological growth, development and otheractivities. The study of nucleic acid is more and more becoming an active field in lifescience. The quantitative determination of nucleic acid is of great importance infundamental research. The study of reaction mechanism between organic smallmolecules and biomacromolecules has very import meaning for finding new drugs、curing diseases form gene levels and studying the reaction mechanism, so it is alwaysscholar’s interesting and attentional subject. Therefore, in this research paper, in orderto analysis the trace components of the nucleic acid and organic dye, RLS methodswere established to the determination of these biological macromolecules of detectingnucleic acid using resonance light scattering. Their experimental conditions have beeninvestigated systematically. The more detailed novelty of this research can becategorized as following：1. The resonance light scattering (RLS) spectra and absorption spectra of theAcridine Orange-yRNA conjugate were studied. The RLS of acridine orange isgreatly enhanced by yRNA in pH11.20. There is a resonance light scattering peak at333nm. Thus, a new RLS method was established for the determination of yRNA.Under optimal conditions, The intensity of RLS signal is proportional to theconcentration of yRNA in the range from0.05~15.0μg/mL with the detection limitof0.017μg/mL. The developed method was applied to the determination of yRNA insynthetic samples with satisfactory results. The method is simple, rapid andsensitivity.2.4-Aminoantipyrine was firstly used to determine yRNA by resonance lightscattering technique. At pH4.78, the interactions of4-Aminoantipyrine with yRNAgive strong RLS signal at472nm. A new method of quantitative determination foryRNA in aqueous solutions has been developed. The linear range for yRNA is0.25~18.0μg/mL, the detection limit is0.23μg/mL for yRNA. The method is simpleoperation, wide linear range, low detection limit. 3. Chapter three is the first report of the interaction of nucleic acid withbromophenol blue(BPB) and cetyltrimethyl ammonium bromide(CTMAB) by RLStechnique. In the buffer solution pH4.78, the RLS of BPB is enhanced a little bynucleic acids. In the system of nucleic acid-CTMAB-BPB, nucleic acid acts as thetemplate to promote the assembly of BPB and surfactant to form nucleicacid-CTMAB-BPB assembled associate. During the formation of associate, CTMABdraws close the distance between BPB and nucleic acids,which enhance theirinteraction and cause enhanced RLS. A new RLS method was established for thedetermination of yRNA. The enhanced RLS intensity at470nm was proportional tothe concentrations of nucleic acids in the range0.15~10.0μg/mL for yRNA and1.0~10.0μg/mL for DNA. The detection limits were0.12μg/mL,0.23μg/mL foryRNA and DNA.