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DNAFingerprinting.ppt

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DNAFingerprintingDNAExtraction:ExtractedDNAmoleculesareincubatedwithrestrictionenzymes(endonucleases).AnEcoR1restrictionenzymeRFLPAnalysis:Molecularbiologistshaveidentifiedregionsofthehumangenomewhererestrictionfragmentlengthsarehighlyvariablebetweenindividuals.VNTRallelesarehighlyvariableregionsofhumanDNA.VNTR’scontinued:Redboxesrepresenttherepeatunitandthebluelollipopsrepresentcutsitesforarestrictionendonuclease.(Here3differentvariants,maybe50inreality).AnalysisofaVNTRlocusmostcommonlyresultsinatwo-bandpattern,onebandinheritedfromeachparent. Aone-bandpatterncanoccurifthesizeofthetwoparentalbandsarethesameornearlythesame.ThepossiblegenotypesareAA,BB,CC,AB,BC,andAC TheRFLPmarkersmostcommonlyusedforDNAprofileanalysisarefoundonchromosomes1,2,4,5,10and17. TheseRFLPmarkersarenamedaftertheirlocationsonthesechromosomes. Forexample,themarkeronchromosome2iscalledD2S44(section44ofchromosome2). ThesechromosomallocationsarealsoreferredtoasDNAloci. TheFederalBureauofInvestigation(FBI)hasbeenaleaderindevelopingDNAtypingtechnologyforuseintheidentificationofperpetratorsofviolentcrime. In1997,theFBIannouncedtheselectionof13STR(shorttandemrepeat)locitoconstitutethecoreoftheUnitedStatesnationaldatabase,CODIS. AllCODISSTRsaretetramericrepeatsequences. AllforensiclaboratoriesthatusetheCODISsystemcancontributetoanationaldatabase. Forexample,D7S280isoneofthe13coreCODISSTRgeneticloci.ThisDNAisfoundonhumanchromosome7. 1aatttttgtattttttttagagacggggtttcaccatgttggtcaggctgactatggagt 61tattttaaggttaatatatataaagggtatgatagaacacttgtcatagtttagaacgaa 121ctaacgatagatagatagatagatagatagatagatagatagatagatagatagacagat 181agatagtttttttttatctcactaaatagtctatagtaaacatttaattaccaatatttg 241gtgcaattctgtcaatgaggataaatgtggaatcgttataattcttaagaatatatattc 301cctctgagtttttgatacctcagattttaaggccDNAprofilesvaryfrompersontoperson.DNAprimershavebeenoptimizedtoallowamplificationofmultipleSTRlociinasinglereactionmixture.Norma’sgenotypeis15,15atthelocusD3S1358,14,16atvWA,and24,25atFGA.ADNAProfile:The13CODISSTRloci Howc