Emulsion PCR A High Efficient Way of PCR Amplification of Random DNA Libraries in Aptamer Selection 英文参考文献.docVIP
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Emulsion PCR A High Efficient Way of PCR Amplification of Random DNA Libraries in Aptamer Selection 英文参考文献
EmulsionPCR:AHighEfficientWayofPCRAmplification
ofRandomDNALibrariesinAptamerSelection
KekeShao1,2,WeifengDing2,FengWang2,HaiquanLi2,DaMa1*,HuiminWang2*
1ClinicalLaboratory,YanchengCityNO.1People’sHospital,Yancheng,China,2LaboratoryCenter,AffiliatedHospitalofNantongUniversity,Nantong,China
Abstract
AptamersareshortRNAorDNAoligonucleotideswhichcanbindwithdifferenttargets.Typically,theyareselectedfroma
largenumberofrandomDNAsequencelibraries.Themainstrategytoobtainaptamersissystematicevolutionofligandsby
exponentialenrichment(SELEX).LowefficiencyisoneofthelimitationsforconventionalPCRamplificationofrandomDNA
sequencelibraryinaptamerselectionbecauseofrelativelowproductsandhighby-productsformationefficiency.Here,we
developedemulsionPCRforaptamerselection.Withthismethod,theby-productsformationdecreasedtremendouslytoan
undetectable level, while the products formation increased significantly. Our results indicated that by-products in
conventional PCR amplification were from primer-product and product-product hybridization. In emulsion PCR, we can
completelyavoidtheproduct-producthybridizationandavoidthemostofprimer-producthybridizationiftheconditions
wereoptimized.Inaddition,italsoshowedthatthemoleculeratiooftemplatetocompartmentwascrucialtoby-product
formation efficiency in emulsion PCR amplification. Furthermore, the concentration of the Taq DNA polymerase in the
emulsionPCRmixturehadasignificantimpactonproductformationefficiency.So,theresultsofourstudyindicatedthat
emulsionPCRcouldimprovetheefficiencyofSELEX.
Citation: Shao K, Ding W, Wang F, Li H, Ma D, et al. (2011) Emulsion PCR: A High Efficient Way of PCR Amplification of Random DNA Libraries in Aptamer
Selection.PLoSONE6(9):e24910.doi:10.1371/journal.pone.0024910
Editor:ReinerAlbertVeitia,InstitutJacquesMonod,France
ReceivedMay1,2011;AcceptedAugust23,2011;PublishedSeptember15,2011
Copyright: ? 2011 Shao et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License,
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