The CRoPS technology combines the power of AFLP as a robust genome complexity reduction method with the power of new sequencing technologies, like the 454 Life Sciences technology highly parallel sequence analysis of DNA fragments.
How it works
A discovery set of individuals/lines/species is selected and their DNA isolated. AFLP fragments are prepared according to the KeyGene AFLP protocol. The KeyGene adaptors are extended with a 4 bp TAG sequence. These DNA fragments are then ligated to the 454/Roche sequence and PCR adaptors for emulsion PCR according to the Roche GS-FLX protocol for library preparation (see diagram).
The fragments are hybridized to the beads and after emulsion PCR sequenced on the GS-FLX. One GS-FLX run generates at least 100Mb of sequence information, resulting in thousands of putative polymorphisms (SSRs, indels and SNPs) of which hundreds will be validated using SNP or SSR detection technologies.
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