Eachkitcontains20individual10-merprimers(suppliedataminimumamountof50nmoleperprimer).TheseprimersaresuitableforuseingeneticmappingandDNAfingerprinting.IntheRAPDtechnique,asingle10merofarbitrarysequenceisusedasaprimerinPCRtoamplifygenomicDNAwherethesequenceoftheDNAiscompletelyunknown.GenomicDNAfromdifferentindividualsgivesdifferentPCRproducts,allowingtheidentificationofDNApolymorphismsthatcanbeusedtoidentifydifferentindividualsorasgeneticMarkers.|ThenumberofPCRproductsgeneratedfromeachgenomicDNAsampledependsontheprimersequence,thegenomicDNAsequence,andthegenomesize.Assumingthattheprimingsitesofa10merRAPDprimerarerandomlydistributedthroughoutagenome,thetheoreticalnumberofPCRproductsisapproximately2.5x10e9xG,whereGisthesizeofthehaploidgenomeinbasepairs.Usingthiscalculation,ahaploidgenomeof2x10e9bp,forexample,isexpectedtogiveabout5PCRproducts.Thispredictionisincloseagreementwithexperimentalresults.
QualityControl:|PurityisverifiedbyanalyticalPAGE.
GCContent
60-70%withoutselfcomplementaryends.
Form:|Suppliedasalyophilizedpowder.
Tm
Asreportedforeachprimer
Mass(ug):t|Asreportedforeachprimer|t|StorageandSt
ABIlity
Storeat-20°C.Lyophilizedpowderisstablefor6monthsat-20°C.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
References
1.Grundmann,etal.,(1997)MulticenterstudyusingstandardizedprotocolsandreagentsforevaluationofreproducibilityafPCR-basedfingerprintingofAcinetobacterspp.J.Clin.MicroBIOLOGyVol.35(12),p3071-3077.2.Berg,D.etal.(1994).MethodsinMolecularandCellularBiology5:13-243.Welsh,J.andM.McClelland(1990).FingerprintinggenomesusingPCRwitharbitraryprimers.NucleicAcidsRes.18(24),7213-72184.5.WilliamsJG,etal.,(1990).DNApolymorphismsamplifiedbyarbitraryprimersareusefulasgeneticmarkers.NucleicAcidsResearch18:6531-65356.Micheli,M.,etal.(1997).InFingerprintingMethodsBasedonArbitrarilyPrimedPCR,pp.55-63.Springer-VerlagPress,NY.7.Graves,LandSwaminathanB(1993).InDiagnosticMolecularMicrobiologyPrinciplesandApplications,pp.617-621.ASMPress,Washington,DC.
