Amplified fragment length polymorphism (AFLP): a review of the procedure and its applications |
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Authors: | M J Blears S A De Grandis H Lee J T Trevors |
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Institution: | (1) Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1, CA;(2) Laboratory Services Division, University of Guelph, Guelph, Ontario, Canada N1G 2W1, CA |
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Abstract: | Amplified fragment length polymorphism (AFLP) is a novel molecular fingerprinting technique that can be applied to DNAs of
any source or complexity. Total genomic DNA is digested using two restriction enzymes. Double-stranded nucleotide adapters
are ligated to the DNA fragments to serve as primer binding 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. Polymorphisms are identified by the presence or absence of DNA fragments following analysis on polyacrylamide
gels. This technique has been extensively used with plant DNA for the development of high-resolution genetic maps and for
the positional cloning of genes of interest. However, its application is rapidly expanding in bacteria and higher eukaryotes
for determining genetic relationships and for epidemiological typing. This review describes the AFLP procedure, and recent,
novel applications in the molecular fingerprinting of DNA from both eukaryotic and prokaryotic organisms.
Received 19 December 1997/ Accepted in revised form 3 June 1998 |
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Keywords: | : AFLP molecular markers genetic mapping PCR polymorphism DNA fingerprinting |
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