AFLP PCR Background
Amplified Fragment Length Polymorphism PCR, also called AFLP PCR was originally described by Zabeau et al., 1993.
AFLP is composed of 3 steps:
1-A) Cellular DNA is digested with one or more restriction enzymes. Typically this involves a combination of two restriction enzymes: a 4 base cutter (MseI) and a 6 base cutter (EcoRI).
1-B) Ligation of linkers (restriction half-site specific adaptors) to all restriction fragments.
2-A) Pre-selective PCR is performed using primers which match the linkers and restriction site specific sequences.
3) Electrophoretic separation and amplicons on a gel matrix, followed by visualisation of the band pattern. The aim of this tool is to perform a theoretical AFLP-PCR experiment by using the same principles, and to suggest the adaptors and primers needed in the experiment.
Applications of AFLP PCR
AFLP is a highly sensitive PCR-based method for detecting polymorphisms in DNA. AFLP can be also used for genotyping individuals for a large number of loci using a minimal number of PCR reactions.
AFLP PCR Protocol
AFLP PCR References
Vos et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research 23: 4407-14.