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Introduction to kinetic (realtime) PCR.
TheDNAFacilityhousesthe“real-time”orkineticPCRinstrument,theAppliedBiosystemsModel7700sequencedetectionsystem(theTaqManinstrument).Thepolymerasechainreaction(PCR)hasrevolutionizedthedetectionofDNAandRNA.Aslittleasasinglecopyofaparticularsequencecanbespecificallyamplifiedanddetected.Theoretically,thereisaquantitativerelationshipbetweenamountofstartingtargetsequenceandamountofPCRproductatanygivencycle.Inpractice,though,itisacommonexperienceforreplicatereactionstoyielddifferentamountsofPCRproduct.Thedevelopmentofreal-timequantitativePCRhaseliminatedthevariABIlitytrADItionallyassociatedwithquantitativePCR,thusallowingtheroutineandreliablequantificationofPCRproducts.Thisinstrument,therefore,nowprovidesinvestigatorswiththeabilitytoperformverysensitive,accurate,andreproducIBLemeasurementsoflevelsofgeneexpression.Inaddition,thisinstrumentcanbeusedinotherapplicationssuchasmeasuringviralload,performingallelicdiscriminationstudies,andoptimizingPCRconditions.
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