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Amplified Fragment Length Polymorphism (AFLP) Analysis on Applied Biosystems Capillary Electrophoresis Systems

Introduction

Amplified fragment length polymorphism (AFLP) analysis is a genetic mapping technique that uses selective amplification of a subset of restriction enzyme-digested DNA fragments to generate a unique fingerprint for a particular genome. First developed for plant studies, AFLP analysis is used for a variety of applications, such as:

Creation of genetic maps for new species.

Determination of relatedness among cultivars

Establishment of linkage groups in crosses

Genetic diversity and molecular phylogeny studiesCreation of genetic maps for new species

Determination of relatedness among cultivars

Establishment of linkage groups in crosses

Genetic diversity and molecular phylogeny studies

The power of AFLP analysis derives from its ability to quickly generate large numbers of marker fragments for any organism, without prior knowledge of the genomic sequence. In addition, AFLP analysis requires only small amounts of starting template and can be used for a variety of genomic DNA samples.

The AFLP procedure consists of two amplification steps: a low-level or preselective amplification, followed by a more selective amplification, which generates a set of fragments that can be used as the discriminatory marker set for a particular sample (Figure 1).

Polymorphisms in band patterns map to specific loci, allowing individuals to be genotyped or differentiated based on the alleles they carry.

Performing Successful AFLP Analysis

The success of an AFLP assay depends on three factors:

1. Optimized reagents

2. A robust and reliable electrophoresis platform

3. Effective analysis software In this Application Note, we will use the Applied Biosyste
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