In consultation with the customer a cDNA-AFLP experimental set-up is chosen and EST's of interest are excised and sequenced.
Its Value
The cDNA-AFLP technology has a wide range of powerful benefits for this application:
the separation of bands on the cDNA-AFLP fingerprints allows the sequencing of different EST's, considerably reducing the redundancy of the EST sequencing required to achieve a reasonable coverage;
cDNA-AFLP fingerprints will allow sequencing of up to 90% of all expressed genes thus resulting in a higher coverage than with conventional EST database construction techniques;
cDNA-AFLP fingerprints of different samples will visualize gene expression differentials that can form the basis of the selection of to be sequenced fragments;
the comparison of a real cDNA-AFLP fingerprint with an in-silico fingerprint of EST's with known sequence allows efficient gene-expression profiling of the already sequenced EST's;
a comparison of an in silico cDNA-AFLP fingerprint of an existing EST database with a real cDNA-AFLP fingerprint can determine the coverage of the EST database and permit the excision and sequencing of new EST's only, allowing very efficient EST database expansion;
AFLP® analyses can also be carried out on a set of cloned EST libraries to assess their genetic diversity in order to allow sequencing of EST libraries with the highest degree of genetic diversity.
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