Evaluation of Efficiency of Different DNA Extraction Protocols and Molecular Identification of Commercial Soybean (Glycine max. , L.) Growing in Turkey

[摘要] FourDNAextractionmethodswereusedtoobtainDNAfromherbariumspecimensofsoybeanandfreshsoybeanseeds,andgerminatedsoybeanseedsinthisstudy.ThequalityofDNAobtainedwasestimatedbyusingaspectrophotometertomeasuretheA260/280absorbanceratio.FourdifferentDNAextractionmethodswerecomparedfortheisolationofDNAfromthesoybeanhomogenates,namelytheCTABextractionmethod,PlantGenomicDNAPurificationKitmethod,EZ1Nucleicacidisolationmethod,andDNAextractionwithphenolpurificationandliquidnitrogen.ThemaingoalofstudyistoevaluatevariousmethodsofDNAisolationintermsofDNAyieldandamplificationquality.TopreserveDNAwell,itisnecessarytodryplantsasfastaspossible.Extractionresultsdependonhowtheplantmaterialisprepared,andthetypeofchemicalsorDNAisolationprotocolsused.ObtaininghighqualityDNAdependsontheisolationtechniqueused.Severalmethodsthatareusefulfordryplanttissuefromherbariumspecimenshavebeendescribed[9,16,19].Fourextractionprotocolswerecomparedusingfreshmaterialanddryherbariumspecimensandseeds.PlantGenomicDNAPurificationKitmethod;theconcentrationofDNAinthesecondelutionextractwasverylow,thusrepeatedelutionwasnotapplied.DNAextractionisdonewithphenolpurificationandliquidnitrogen.Thistechniqueisnotquitesuitableforfreshplantleaves.Itisusuallyusedforherbariumsamplesofatleast0.2gCTABextractionmethod.ThisprotocolisquitesuitableforDNAisolationfromthefreshplantmaterialorgerminatedseedplantlets.ThetotalDNAwasisolatedfromapproximately0.5-1goffreshleaves(conservedinCTAB)ofthesamecollectionofsample.EZ1NucleicacidisolationtehniqueisquiteusefulforhighyieldandqualityofDNAisolationfromdriedsoybeanseeds.Inthismethods,nofurtherpurificationwasneededformolecularanalysis.

[学科分类] 生物科学（综合）

[关键词] Genomic DNA extraction;soybean (Glycine max. L.)