Eachkitcontains20individual10-merprimers(suppliedataminimumamountof50nmoleperprimer).TheseprimersaresuitableforuseingeneticmappingandDNAfingerprinting.IntheRAPDtechnique,asingle10merofarbitrarysequenceisusedasaprimerinPCRtoamplifygenomicDNAwherethesequenceoftheDNAiscompletelyunknown.GenomicDNAfromdifferentindividualsgivesdifferentPCRproducts,allowingtheidentificationofDNApolymorphismsthatcanbeusedtoidentifydifferentindividualsorasgeneticMarkers.|ThenumberofPCRproductsgeneratedfromeachgenomicDNAsampledependsontheprimersequence,thegenomicDNAsequence,andthegenomesize.Assumingthattheprimingsitesofa10merRAPDprimerarerandomlydistributedthroughoutagenome,thetheoreticalnumberofPCRproductsisapproximately2.5x10e9xG,whereGisthesizeofthehaploidgenomeinbasepairs.Usingthiscalculation,ahaploidgenomeof2x10e9bp,forexample,isexpectedtogiveabout5PCRproducts.Thispredictionisincloseagreementwithexperimentalresults.
Vial-01:5’-CAGGCCCTTC-3’|Vial-02:5’-TGCCGAGCTG-3’|Vial-03:5’-AGTCAGCCAC-3’|Vial-04:5’-
AATCGGGCTG-3’|Vial-05:5’-AGGGCTCTTG-3’|Vial-06:5’-GGTCCCTGAC-3’|Vial-07:5’-GAAACGGGTG-3’|Vial-08:5’-GTGACGTAGG-3’|Vial-09:5’-GGGTAACGCC-3’|Vial-10:5’-GTGATCGCAG-3’|Vial-11:5’-CAATCGCCGT-3’|Vial-12:5’-TCGGCGATAG-3’|Vial-13:5’-CAGCACCCAC-3’|Vial-14:5’-TCTGTGCTGG-3’t|Vial-15:5’-TTCCGAACCC-3’|Vial-16:5’-AGCCAGCGAA-3’|Vial-17:5’-GACCGCTTGT-3’|Vial-18:5’-AGGTGACCGT-3’|Vial-19:5’-CAAACGTCGG-3’|Vial-20:5’-GTTGCGATCC-3’
QualityControl:PurityisverifiedbyanalyticalPAGE.
GCContent:60-70%withoutselfcomplementaryends.
MolecularWeight:Asreportedforeachprimer
Storeat-20°C.Lyophilizedpowderisstablefor6monthsat-20°C.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
References
1.Grundmann,etal.,J.Clin.MicroBIOLOGy35(12):3071-3077(1997).2.Berg,D.,etal.,MethodsinMolecularandCellularBiology5:13-24(1994).3.Welsh,J.andMcClelland,M.,NucleicAcidsRes.18(24):7213-7218(1990).4.Williams,J.G.,etal.,NucleicAcidsResearch18:6531-6535(1990).5.Micheli,M.,etal.,(1997).InFingerprintingMethodsBasedonArbitrarilyPrimedPCR,pp.55-63.Springer-VerlagPress,NY.6.Graves,LandSwaminathan,B.,(1993).InDiagnosticMolecularMicrobiologyPrinciplesandApplications,pp.617-621.ASMPress,Washington,DC.
