modulation of non-templated nucleotide addition by taq dna polymerase primer modifications

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1、ShortTechnicalReportsSHORTTECHNICALModulationofNon-Tem-cer,datacollectionandallelesizingarebasedontheuseofGENESCANÔandREPORTSplatedNucleotideAdditionGENOTYPERÔsoftware.Atpresent,ManuscriptspublishedintheShortbyTaqDNAPolymerase:manualeditingisrequiredtocorrectTechnicalReportssectionareshorterPri

2、merModificationsthatallelemisidentificationduetovariabili-andlesscomprehensiveinscopethantyinnon-templatedadditionofasinglefullResearchReports.FacilitateGenotypingnucleotide,mainlyadenosine,tothe3¢endofthefluorescentlylabeledstrandBioTechniques20:1004-1010(June1996)byTaqDNApolymerase(1).Recentl

3、y,weshowedthatthedegreeABSTRACTtowhichamarkerissubjectto“plusA”modificationisprimer-specific(7),butTaqDNApolymerasecancatalyzenon-wedidnotdefinethestructuralmotif(s)templatedadditionofanucleotide(princi-underlyingthisspecificity.Instead,wepallyadenosine)tothe3¢endofPCR-am-demonstratedthatgenoty

4、pingerrorsplifiedproducts.Recently,weshowedthatduetovariabilityinadenosineadditionthisactivity,whichisprimer-specific,pre-couldbeminimizedbyamplifyingsentsapotentialsourceoferroringenotyp-markersinoneoftwothermalcyclingingstudiesbasedontheuseofshorttandemprotocols:onethatfavorsproductionofrepea

5、t(STR)markers.Furthermore,inre-the“true”alleleandanotherthatfavorsviewingourdata,wefoundthatnon-tem-theproductionofthe“plusA”product.platednucleotideadditionadjacenttoa3¢Inthecourseofthesestudies,wedis-terminalCisfavoredandthatadditionad-coveredasmallnumberofmarkersjacenttoa3¢terminalAisnot.Itw

6、asclear,highlyresistanttoadenylationandhowever,thatfeaturesofthetemplateinad-someadditionalmarkersthatgaveditiontothe3¢terminalbasealsoaffecttheadenylatedproductsevenwhenthefractionofproductadenylated.TodefinePCRconditionsusedfavoredproduc-consensussequencesthatpromoteorinhib-tionofthetrueallel

7、e.Wedecidedtoitproductadenylation,wetransplantedse-studythesemarkersinhopesofcreat-quencesbetweenthe5¢endsofthereverseingnewmarkerpanelswiththefollow-primersofmarkersthatareadenylatedandingfeatures:(i)uniform(allornone)thoseofmark