吡啰红Y共振光散射法测定纳克级核酸.docVIP

吡啰红Y共振光散射法测定纳克级核酸 第18卷第3期 2OO4年9月 南华大学(自然科学版) JournalofNanhuaUniversity(ScienceandTechnology V01.18No.3 Scp.2OO4 ArticleID:1673——0062(2004)03——0012——06 DeterminationofNucleicAcidsatNanogramLevelswith PyronineYbyaResonanceLight——ScatteringTechnique WANGYong—sheng,XIAOXi—lin,LIGui—rong,LUChang一 (1.SchoolofPublicHealth,NanhuaUniversity,Hengyang,Hunan421001,China;2.Schoolof ChemistryandChemicalEngineering,NanhuaUniversity,Hengyang,Hunan421001,China) Abstract:Basedontheenhancementofresonancelightscattering(RLS)ofPyronineY(PY) bynucleicacids,anewquantitativedeterminationmethodfornucleicacidsinthenanogram rangehasbeendeveloped.InthepHrangel1.5—12.0.theRLSofPYisgreatlyenhancedby nucleicacids,withthemaxilnulnscatteringpeaklocatedat364.0nm.eenhancedintensityof RLS,?I吣,wasfoundtobeproportionaltotheconcentrationofnucleicacidsintherange 27.0,625ng/mLforfishspermDNA.39.0,500ng/mLforcalfthymusDNAand59.0, 375ng/mLforyeastRNA.Thelimitsofdeterminationwere8.1ng/mLforfishspermDNA, l1.7ng/mLforcalfthymusDNAand17.7ngCmLforyeastRNA.Sixsyntheticsampleswere determinedsatisfactorily.I11eassayisconvenient.rapidandinexpensive. Keywords:pyronineY;nucleicacids;resonancelightscattering(RLS) CInumber:o657.32Documentcode:B 吡红Y共振光散射法测定纳克级核酸 王永生,肖锡林,李贵荣,吕昌银 (1.南华大学公共卫生学院,湖南衡阳421001;2.南华大学化学化工学院,湖南衡阳 421001) 摘要:研究了二苯并吡喃类染料吡口罗红Y(PY)与核酸作用的共振光散射光谱,在 pill1.5,12.0s—NaOH缓冲溶液中,1.75×10mol/L的PY溶液加入核酸后,体 系的共振光散射增强,在364.0nJffl处,存在一共振光散射增强峰,其增强强度(?I 吣 = I—Io)与核酸浓度呈线性关系,据此建立了一种测定纳克级核酸的简便灵敏的方 法.对于fsDNA,方法的线性范围为27.0,625ng~mL,检出限为8.1ng~mL;对于ctD — NA,方法的线性范围为39.0,500ng/mL,检出限为l1.7ng~mL;对于yRNA,方法的 线 性范围为59.0,375ng/mL,检出限为17.7ng/mL.建立的方法已用于六种合成样品 的 测定,结果令人满意. 关键词:吡口罗红Y;核酸;共振光散射法 中图分类号:0657.32文献标识码:B Receiveddate:2004一o4—19 Foundationitem:theSciencesResearchProgramofEducationalDepartmentofHunanProvi nce(01C213) Biography:WANGYong—sheng(1950一),male,bominChang~ing,Hunan,Professor. 第l8卷第3期王永生等:吡红Y共振光散射法测定纳克级核酸l3 1Introduction Thequantitativeanalysisofnucleicacidsisof greatinterestsinceitiso