东北红豆杉RAPD反应体系的优化(摘要)(英文)

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1、AgriculturalScience＆Technology，2010，11(3)：47—49Copyright⑥2010，InformationInstituteofHAAS．AllrightsresemedAgriculturalBiotechnologyOptimizationofRAPDReactionSystemofTaxuscuspidateSieb．etZuccXlAOLu，XUMing-zi，CUIMiao，LIUBoDepartmentofAgriculture，AgriculturalCollegeofYanbianUniversity，Longjing133400

2、Abstract『0biective]TheresearchaimedtostudytheoptimizationconditionsofRAPDreactionsystemofTaxuscuspidata．『Method]UsingtheIeavesofTaxuscuspidataasmaterials．DNAwasextractedbyusingmodifiedCTABmethod．TheinfluencesoftemplateDNAconcentration．primerconcentration．dNTPconcentration．qDNApolymeraseamountand

3、magnesiumionconcentrationontheamplificationefectsofRAPDofTaxuscuspidatewereanalyzed．『ResultlThroughanalysisandcomparisonofvariousfactors，theoptimizedreactionsystemwasestablishedasfollows：inatotalvolumeof20pl，containing1OngDNA，2．0mmol／LMg“，O．2mmol／LdNTPs，15pmol／plrandomprimer，1．OUTaqpol—ymerase，2

4、uI10xbufer．andthesupplementaryddHO．IConclusionTheresearchprovidedtechnicaIsupportforfurtherdiscussiononthere—IatedstudieswiththesexualidentificationofTaxuscuspidate．KeywordsTED(UScuspidata；RAPD；ReactionsystemTaxuscuspidateSieb．etZuccknownastaxusorChineseat一2O℃wasalsomixedtogethertoputintorefrige

5、ratoratyew．belongstoTa)(aceaeS．F．Grey．Taxales．Coniferop·一2OoCfor30minuntiItherewereflocculentprecipitates．sudec，Gymnospermae，subphylumofgymnosperml1l_TaxolisNext．sampleswerecentrifugedfor10minwith11O00r／minakindofditerpenoidsintaxusandprimarilyobtainedfrOmTaxustodischargethesupernatant，andthenfl

6、ushedorprecipitatedbrevifoliabyWanii2．wh_leitsspeciaIanticancermechanismhaswith70％alcohoIforthreetimestoinvertcentrifugetubesona~ractedpeople’swideentions．Taxolisadioeciousplant．theblottingpaperfordrying．(1xTEbuferwasaddedintoandcurrentlythereisnealygender—relatedstudy．Withfemalesamplestodissolv

7、eDNA．and2ulRnasewasalsomixedto—plantIeavesofTaxuscuspidateasthematerials．RAPDreac—aethertoremoveRNAbyincubationfor1hat37℃．(6)0．3tionconditionswereoptimizedInthisstudy-6j．whichaimedmol／LNaCIandequalvolumeofanhydrousethano1wer