Autors:

CiteWeb id: 19950000004

CiteWeb score: 21767

DOI: 10.1093/nar/23.21.4407

A novelDNAfingerprinting technique called AFLPis described. TheAFLPtechnique isbasedon the selective PCRamplification ofrestriction fragments fromatotal digest ofgenomicDNA.Thetechnique involves threesteps: (i) restriction oftheDNA and ligation ofoligonucleotide adapters, (ii) selective amplification ofsetsofrestriction fragments, and(iii) gelanalysis oftheamplified fragments. PCRamplification ofrestriction fragments isachieved byusing the adapter andrestriction site sequence astarget sites for primerannealing. The selective amplification is achieved bytheuseofprimers thatextend into the restriction fragments, amplifying onlythose fragments inwhichtheprimer extensions matchthenucleotides flanking therestriction sites. Usingthis method, sets ofrestriction fragments maybevisualized byPCR without knowledge ofnucleotide sequence. The methodallows thespecific co-amplification ofhigh numbersofrestriction fragments. Thenumberof fragments that canbeanalyzed simultaneously, however, isdependent ontheresolution ofthedetection system. Typically 50-100 restriction fragments are amplified anddetected ondenaturing polyacrylamide gels. TheAFLPtechnique provides anovel andvery powerful DNAfingerprinting technique forDNAsofany origin orcomplexity.

Links: