The KeyMaps technology is a new application of the AFLP® technology that allows the construction and elaboration of dense integrated genetic and physical maps.
How it works
The crux of the KeyMaps concept is the in silico linkage of genetically mapped (AFLP) markers with AFLP fragments found in BAC-pools and individual BACs. The AFLP fingerprints are analysed by capillary electrophoresis on the MegaBace 1000 capillary sequencer. This allows highly accurate fragment sizing. The physical map construction is based on matching the +0/+0 AFLP fingerprints of all individual BACs. Depending on the coverage of the BAC-library, the matching of individual BAC-fingerprints can result in a physical map with a density of one AFLP marker per 40 kb.
The physical map is then linked to the genetic map by fingerprinting BAC-pools with the +2/+3 primer combinations of the genetic markers of interest after which the BAC-pools and the individual BACs with the marker of interest are identified. Depending on the overlap of the BACs in which the marker has been found, the marker can be mapped on a specific region of these BACs. The number of BAC-pools required for the linkage of the physical and genetic map depends on the number of BAC clones and the total genome size but usually the number of fingerprints required does not exceed one AFLP gel of 96 fingerprints.
Once all individual BACs have been fingerprinted, the KeyMaps software will allow the physical mapping of any future marker of interest. Fingerprinting the BAC-pools with the +2/+3 combination of interest and (reasonably) accurate fragment size information is all that it is required.
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