【文档说明】PCR技术在医学中的应用-课件.ppt，共(43)页，12.447 MB，由小橙橙上传

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PCR技术在医学中的应用WhatisPCRHowtoPCRWhydoPCRDevelopedin1983byKaryMullispolymerasechainreaction(PCR)1993年的诺贝尔化学奖AnimationofPCRPCRReagentsTempl

ateDNAPrimersdNTPDNAPolymeraseBufferMg2+QuestionsAnneallingDNApolymerasev.s.WhyprimersareessentialforDNApolymerase?In

itializationstepDenaturationstepAnnealingstepElongationstepFinalelongationFinalholdPCRprocedure&stages20~40CyclesExponentialamplif

icationLevelingoffstagePlateauEvolutionofmachinesforPCR1234PCROptimizationContaminationHairpinsPolymeraseerrorsM

agnesiumconcentrationNon-specificprimingPrimerdimersDeoxynucleotidesGC-richTemplateProofreadingDNAPolymerasePfu,KOD…DMSO,betai

neRedesignPrimersHot-StartPolymerasePCROptimization:PrimerDesignSoftwaresPrimerpremier:e-PCR(NCBI)Oligo7:Primer3:mandlinestyledesigntoolMulti-

OligodesignforprobesorprimersSinglepairedprimersdesignforPCRAvoidnon-specificamplificationbyblastPrimersforamplific

ationofhumanb-actingenomicDNAregionPrimerDesign:avoidfailureofamplification5’-primer:stem-loopstructurew

ithhighmeltingtemperatureHighdifferencebetweenproductandprimermeltingtemperatures.3'-endForwardPrimerdimer.PrimerDesign:avoidfailureofamplif

ication3'-enddimerbetweentheForwardandReverseprimers.TerminalstabilityoftheForwardPrimeristoohigh.PrimerDesign:avoidfailureof

amplificationPCRmethodsIsothermalPCRReverseTranscriptionPCRFrequentlyusedPCRmethodsReal-TimePCR(quant

itativePCR)Overlap-extensionPCRNestedPCRFrequentlyusedPCRmethodsReverseTranscriptionPCROverlap-extensionPCRFrequentlyusedPCRmethodsNeste

dPCRFrequentlyusedPCRmethodsFrequentlyusedPCRmethodsReal-TimePCR(quantitativePCR)DetectfluorescenceforeachcycleQuantitaivebyamplifica

tioncurveQuanlitycontrolReal-TimePCRdouble-strandedDNA-bindingdyeReal-TimePCR(quantitativePCR)DetectfluorescenceforeachcycleReal-TimePCR(q

uantitativePCR)QuanlitycontrolbymeltingcurveFluorescentreporterprobeReal-TimePCRReal-TimePCRPathogenDiagnosismicroRNADiagnosisheredopa

thiaDiagnosisIso-ThermalPCRIso-ThermalFastSensitiveLAMP/RPALoop-mediatedisothermalamplification(LAMP)Rebinasepolymeraseamplificat

ionTwoprimersQuantitativableSensitiveLongPrimersApplicationofPCRApplicationofPCRSelectiveDNAisolationAmplificationa

ndQuantificationPCRindiagnosisofdiseasesPCRinforensicsciencePCRinforensicsciencevariablenumberoftandemrepeatsMitochondriaSequencingShortTandemR

epeatMitochondriaSequencingPCRinforensicscienceShortTandemRepeatvariablenumberoftandemrepeatsPCRinforensicscienceSexdiscernmen

tY-chromosomespecificDNAMedicalapplicationofPCRMedicalapplicationsGenetictestingTissuetypingTumortargettypingCellFreeDNAan

alysismajorcurrenttechniquesforanalyzingfetalchromosomesPCRindiagnosisoffetalheredopathiaBabygenderbloodtestsFetalsexdiscernme

ntChorionicvillussamplingObstetricultrasonographyCell-freefetalDNAafterthe13thweekofpregnancyAccuracy:~90%riskofdamageto

thefetusafterthe11th~15thweekofpregnancyafterthe7thweekofpregnancyAccuracy:~98%FetalsexdiscernmentProc

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