title: Amplified fragment-length polymorphism analysis.
abstract: Amplified fragment length polymorphism (AFLP) analysis is a universal polymerase chain reaction (PCR)-based DNA fingerprinting technique comprising three main stages: (i) digestion of genomic DNA with restriction endonucleases and ligation to double-stranded adaptors (each comprised of two oligonucleotides), thus creating restriction fragments with identical known adaptor sequences; (ii) specific amplification of a subset of these DNA fragments using primers (one labeled) targeting the adaptor sequences and additional selected bases within the unknown genomic DNA; and (iii) analysis of the patterns (usually automated). Differences or polymorphisms between samples are revealed by separation of the labeled fragments by electrophoresis (standard agarose, high-resolution denaturing acrylamide, or capillary gels). Comparison of banding patterns is typically achieved using dedicated fingerprinting analysis software. The advantages of AFLP analysis include the ability to use a universal protocol in combination with different restriction endonucleases and the choice of adding one or more selective nucleotides in the PCR -primers to achieve optimal results relatively quickly without prior knowledge of DNA sequences from a large variety of (micro)organisms. The method also has the potential for high-throughput and local electronic database pattern storage with relatively low cost. Disadvantages include variation in the precision of sizing of fragments, leading to suboptimal reproducibility, particularly across different platforms.
authors.affiliation: Respiratory and Systemic Infection Laboratory, Health Protection Agency, Centre for Infections, London, UK.
authors.forename: Norman K
authors.initials: NK
authors.lastname: Fry
authors.forename: Paul H M
authors.initials: PH
authors.lastname: Savelkoul
authors.forename: Paolo
authors.initials: P
authors.lastname: Visca
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis / Q000379:methods* / Q000706:statistics & numerical data; D016172:DNA Fingerprinting / Q000379:methods; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005456:Fluorescent Dyes; D005827:Genetics, Microbial / Q000379:methods*; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification; D017720:Molecular Epidemiology / Q000379:methods*; D010761:Phosphorus Radioisotopes; D012984:Software
chemical_list: D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D010761:Phosphorus Radioisotopes; D004262:DNA Restriction Enzymes