The Oligo-FISH (Fluorescent in situ hybridization) probes were designed based on the synteny of Lens culinaris CDC Redberry genome (Lcu.2RBY), against the wild Lens genome assemblies aiming to confirm most of the rearrangements between those genomes physically (using bedtools instersect). Probe locations in Lcu.2RBY were selected to maximize the number of rearrangements that could be confirmed against all wild Lens genomes. Arbor Biosciences designed and synthesized the probes using Lcu.2RBY. The genome assembly was divided into oligo-sequences of at least 45 nucleotides in length. These sequences were filtered for desired location, specificity, melting temperature, and sequence conservation relative to the wild Lens assemblies (using BlastN). Two different oligo pools were generated and labeled with different colours – green and red. The in silico location of these probes in each Lens species assembly (peak enrichment) was used to predict the physical (in situ) location of the probes during FISH. The combination of the oligo-pools/colours gives a specific pattern (also called barcode) for each Lcu.2RBY and wild Lens chromosomes. When this pattern differs from the reference (Lcu.2RBY), it confirms a rearrangement in the compared wild species.