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1、JournalofForestryResearch,17(1):31–34(2006)31ComparisonsofextractionandpurificationmethodsofsoilmicroorganismDNAfromrhizospheresoil1,2232JIAXia,HANShi-jie,ZHAOYong-hua,ZHOUYu-mei1EnvironmentalScienceandEngineeringCollege,Chang’anUniversity,Xi’an710054,P.R.China2InstituteofAppliedEcologyof
2、theChineseAcademyofSciences,Shenyang110016,P.R.China3CollegeofEarthScienceandLandResourceManagement,Chang’anUniversity,Xi’an710054,P.R.ChinaAbstract:MicroorganismDNAofrhizospheresoilfromPinuskoraiensisandPinussylvestriformiswereextractedbyproteinaseKbasedonSDSmethod,CTABmethod,PVP(polyvin
3、ylpolypyrrolidone)method,andfreezingandthawingmethodandthecrudeDNAfromrhizospheresoilwerepurifiedbydialysismethod,silverbeadsabsorptionmethod,andsqueezingDNAgelmethod.Theresultsofdifferentextractingandpurifyingmethodswerecomparedandevaluated.Resultsindicatedthatthebestmethodofextractionfo
4、rmicroorganismDNAinrhizospheresoilwasproteinseKbasedonSDSmethodwithhighsaltconcentrationof1.0%(w/v)NaCl,whichcouldeffectivelyeliminatehumicacidsandotherimpurities.ThedialysismethodwassuitabletopurifyDNAfromrhizospheresoilbecauseofeffectivelyremovingbrownmattersandhumicacidsandthepurifiedp
5、roductsweresuitedtoPCRamplification.SqueezingDNAgelmethodwasalsoagoodpurificationmethodwiththeadvantageofinexpensiveincostandefficientinuse.Keywords:Extraction;MicroorganismDNA;Pinuskoraiensis;Pinussylvestriformis;Purification;RhizospheresoilCLCnumber:S154.3Documentcode:AArticleID:1007-66
6、2X(2006)01-0031-04Introductionnationsofplentyofhigh-moleculehumicacids,exudationsfromroots,andotherhigh-moleculecompoundsdecomposedfromMoleculetechniquesbasedonnucleicacidhaveledtotre-plantrootresiduesareverycrucialproblemsattheprocessesofmendouschangesindetectingnaturalenvironmentalmicro
7、or-PCRamplification(Smallaetal.1993).Therefore,theselectionganism(Liesacketal.1992;Wardetal.1990).Inthepast,aofextractionandpurificationmethodsisaverycrucialforDNAcultivationdependentapproachonlycanexamine0.1%－1%ofofrhizospheresoilmicroorganism.totalenvironmentalmic
