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1、■ORIGINALARTICLE■COMPARISONOFWHOLEGENOMEAMPLIFICATIONMETHODSFORFURTHERQUANTITATIVEANALYSISWITHMICROARRAY-BASEDCOMPARATIVEGENOMICHYBRIDIZATIONYun-ShienLee1,2,Chi-NeuTsai3,Chia-LungTsai4,Shuenn-DyhChang4,Ding-WeiHsueh1,Chun-TingLiu1,Chung-ChunMa1,Seng-HsuanLin1,Tzu-HaoWang1,4*,Hsin-ShihWang3,41Genomic
2、MedicineResearchCoreLaboratory,ChangGungMemorialHospital,2DepartmentofBiotechnology,Ming-ChuanUniversity,3GraduateInstituteofClinicalMedicalSciences,CollegeofMedicine,and4DepartmentofObstetricsandGynecology,ChangGungMemorialHospital,ChangGungUniversity,Tao-Yuan,Taiwan.SUMMARYObjective:Wholegenomeamp
3、lification(WGA)isacrucialprocedureforgenomicDNAanalysisfromlimitedsources,suchasinforensicanalysis,embryobiopsyforpreimplantationgeneticdiagnosis,orneedleaspirationbiopsies.SeveralstrategiesforWGAhavebeendevelopedforeithergenotypingormicroarray-basedcomparativegenomehybridization(array-CGH)duringthe
4、pastdecade.Nevertheless,therewerefewstudiesinwhichvariousWGAmethodshadbeenperformedside-by-sideandresultsevaluatedwithmultiplemethods.MaterialsandMethods:Easeofperformance,qualitativeaccuracy,andquantitativefidelityofdifferentWGAmethods,suchasdegenerateoligonucleotide-primedpolymerasechainreaction(D
5、OP-PCR),ligation-mediatedPCR(LM-PCR)andstranddisplacementamplification(SDA),werecomparedinamplifyinggenomicDNAderivedfromkaryotype-confirmedamniocytesandthecancercelllineSAOS2.Results:Usinganalysiswithmicrosatellitemarkers,singlenucleotidepolymorphismmarkers,andarray-CGH,ourresultssuggestedthat:(1)g
6、enomicDNAamplifiedfromDOP-PCRresultedinfalsepositiveandnegativeresultsbyanalysiswitharray-CGH;(2)SDAistheeasiestperformancemethodamongthethreeWGAmethods;and(3)amplifiedDNAproductsgeneratedbyLM-PCRbestreflecttheoriginalgenomicDNA.Conclusion:TheamplifiedDNAproductsgeneratedbyLM-PCRbestreflecttheorigin
7、algenomicDNA.[TaiwanJObstetGynecol2008;47(1):32–41]KeyWords:array-CGH,DOP-PCR,LM-PCR,SDA,wholegenomeamplificationIntroductionisagenome-widedetectingmethodforimbalancedcopynumberofDNAsequences[2–4].Chr
