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556<br />

– Low intensity of the bands: insufficient amount of primer or dNTPs. Try<br />

increasing the amount of each one of these components at a time.<br />

3 AFLP<br />

Emanuele G. Biondi<br />

Principle<br />

Amplified Fragment Length Polymorphism (AFLP) is a recently developed<br />

technique based on restriction and amplification (Zabeau et al, 1993; Vos et al.<br />

1995; Fig. 2). Using this method it is possible to generate up to 100 genomic<br />

markers with a single combination of restriction enzyme and primers. In particular,<br />

the application to microbial population analysis has been used to differentiate<br />

bacteria at strain and species levels from the taxonomic, phylogenetic<br />

or the population genetics point of view (Biondi et al. 2003). Moreover,<br />

adapter<br />

GAATT<br />

C<br />

GAATT<br />

C<br />

primer<br />

genomic DNA<br />

EcoRI MseI<br />

GAATTC<br />

CTTAAG<br />

C<br />

TTAAG<br />

C<br />

TTAAG<br />

GAATTC<br />

CTTAAG<br />

AATT<br />

TTAA<br />

Digestion with EcoRI and MseI<br />

T<br />

AAT<br />

Ligation with Adapters<br />

T<br />

AAT<br />

TTAA<br />

AATT<br />

TAA<br />

T<br />

TAA<br />

T<br />

Annealing of Selective primers<br />

adapter adapter<br />

Amplification<br />

primer<br />

adapter<br />

Fig. 2. Outline of AFLP technique. In dark grey the EcoRI restriction site and in light<br />

grey the MseI restriction site. See text for details

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