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核酸恒温扩增技术(HDA).doc

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EMBORep.2004August;5(8):795–800. Publishedonline2004July9.doi:10.1038/sj.embor.7400200.PMCID:PMC1249482HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/about/copyright.html"Copyright©2004,EuropeanMolecularBiologyOrganization ScientificReport Helicase-dependentisothermalDNAamplification MyriamVincent,1*YanXu,1*andHuiminKong1a 1NewEnglandBiolabs,32TozerRoad,Beverly,Massachusetts01915,USA aTel:+19789275054;Fax:+19789211350;E-mail:HYPERLINK"mailto:kong@neb.com"kong@neb.com *Theseauthorscontributedequallytothiswork ReceivedJanuary14,2004;RevisedMay24,2004;AcceptedJune14,2004. ThisarticlehasbeenHYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/citedby/"citedbyotherarticlesinPMC. HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"#"OtherSections▼ HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"#"Abstract HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"__secid605336#__secid605336"Introduction HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"__secid133855#__secid133855"Results HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"__secid467122#__secid467122"Discussion HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"__secid164112#__secid164112"Methods HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"S1#S1"SupplementaryMaterial HYPERLINK"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1249482/"\l"__ref-listid121790#__ref-listid121790"References Abstract PolymerasechainreactionisthemostwidelyusedmethodforinvitroDNAamplification.However,itrequiresthermocyclingtoseparatetwoDNAstrands.Invivo,DNAisreplicatedbyDNApolymeraseswithvariousaccessoryproteins,includingaDNAhelicasethatactstoseparateduplexDNA.WehavedevisedanewinvitroisothermalDNAamplificationmethodbymimickingthisinvivomechanism.Helicase-dependentamplification(HDA)utilizesaDNAhelicasetogeneratesingle-strandedtemplatesforprimerhybridizationandsubsequentprimerextensionbyaDNApolymer