Amplified fragment length polymorphism (AFLP): a review of the procedure and its applications

Amplified fragment length polymorphism (AFLP) is a novel molecular fingerpr inting technique that can be applied to DNAs of any source or complexity. T otal genomic DNA is digested using two restriction enzymes. Double-stranded nucleotide adapters are ligated to the DNA fragments to serve as primer bi nding sites for PCR amplification. Primers complementary to the adapter and restriction site sequence, with additional nucleotides at the 3'-end, are used as selective agents to amplify a subset of ligated fragments. Polymorp hisms are identified by the presence or absence of DNA fragments following analysis on polyacrylamide gels. This technique has been extensively used w ith plant DNA for the development of high-resolution genetic maps and for t he positional cloning of genes of interest. However, its application is rap idly expanding in bacteria and higher eukaryotes for determining genetic re lationships and for epidemiological typing. This review describes the AFLP procedure, and recent, novel applications in the molecular fingerprinting o f DNA from both eukaryotic and prokaryotic organisms.