南京林业大学学报（自然科学版）

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2000, Vol. 24 ›› Issue (01): 27-31.doi: 10.3969/j.jssn.1000-2006.2000.01.007

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Optimization of RAPDAmplification Programin Thin-walled Microcentrifuge TubeswithIdaho Rapidcycler

GanSiming ShiJisen (LaboratoryofForestGeneticsandGeneEngineeringNanjingForestryUniversity Nanjing 2 10 0 37) BaiJiayu   

  1. ResearchInstituteofTropicalForestryChineseAcademyofForestry
  • Online:2000-02-18 Published:2000-02-18

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Abstract: Inthin walledmicrocentrifugetubes,optimizationofRAPDamplificationprogramforEucalyptusurophyl laS .T .BlakewasmadewithIdahoRapidcycler.Thetemperatureandtimefordenaturation ,annealingandelongation aswellasthenumberofcycleswereindividuallyoptimized .Theprogramachievedwasasfollows :predenaturationat 93℃for1min ;then 40cycles:denaturation 2 0sat93℃ ,annealing2 1sat38℃ ,andelongation 1minat72℃ ; andatlastholding 7minat72℃ .TheprogramcouldresultinsatisfactoryspecificityandrepeatabilityinRAPD amplificationinrelativelyshorttime.Inaddition ,theinnerrowofreactionsinthin walled tubestandperformedweak oremptyinamplificationoflongfragments ,soonlytheouterrowcouldbeusedinpracticaloperation .

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