Eachkitcontains20individual10-merprimers(suppliedataminimumamountof50nmoleperprimer).TheseprimersaresuitableforuseingeneticmappingandDNAfingerprinting.IntheRAPDtechnique,asingle10merofarbitrarysequenceisusedasaprimerinPCRtoamplifygenomicDNAwherethesequenceoftheDNAiscompletelyunknown.GenomicDNAfromdifferentindividualsgivesdifferentPCRproducts,allowingtheidentificationofDNApolymorphismsthatcanbeusedtoidentifydifferentindividualsorasgeneticMarkers.|ThenumberofPCRproductsgeneratedfromeachgenomicDNAsampledependsontheprimersequence,thegenomicDNAsequence,andthegenomesize.Assumingthattheprimingsitesofa10merRAPDprimerarerandomlydistributedthroughoutagenome,thetheoreticalnumberofPCRproductsisapproximately2.5x10e9xG,whereGisthesizeofthehaploidgenomeinbasepairs.Usingthiscalculation,ahaploidgenomeof2x10e9bp,forexample,isexpectedtogiveabout5PCRproducts.Thispredictionisincloseagreementwithexperimentalresults.
Storeat-20°C.Lyophilizedpowderisstablefor6monthsat-20°C.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
References
1.Grundmann,Towner,Dijkshoorn,Gerner-Smidt,Maher,SeifertandVaneechoutte.(1997)MulticenterstudyusingstandardizedprotocolsandreagentsforevaluationofreproducibilityafPCR-basedfingerprintingofAcinetobacterspp.J.Clin.MicroBIOLOGyVol.35(12),p3071-3077.2.Berg,D.etal.(1994).MethodsinMolecularandCellularBiology5:13-24.3.Welsh,J.andM.McClelland(1990).FingerprintinggenomesusingPCRwitharbitraryprimers.NucleicAcidsRes.18(24),7213-7218.4.WilliamsJG,KubelikAR,LivakKJ,RafalskiJA,TingeySV(1990).DNApolymorphismsamplifiedbyarbitraryprimersareusefulasgeneticmarkers.NucleicAcidsResearch18:6531-6535.5.Micheli,M.,etal.(1997)InFingerprintingMethodsBasedonArbitrarilyPrimedPCR,pp.55-63.6.Springer-VerlagPress,NY.Graves,LandSwaminathanB(1993).InDiagnosticMolecularMicrobiologyPrinciplesandApplications,pp.617-621.ASMPress,Washington,DC.
