We present a 12.5x sequence-based physical map of the cultivated sunflower genome based on a whole-genome profiling (WGP) technique developed by Keygene NV. Fourteen-genome equivalent BAC libraries were constructed and pooled two-dimensionally, and the restriction fragments from the pooled BAC clones were sequenced using the Illumina Genome Analyzer platform. WGP takes advantage of these short read sequences to generate 20-30 unique sequence tags for each BAC clone. BAC pools were tagged individually and based on the coordinates in the 2D pool screening, unique sequence tags were assigned to individual BACs. The BAC clones were then ordered into contigs based on shared regions containing identical sequence tags, using a modified version of Finger Print Contigs (FPC) -the software that has been routinely used for restriction profile-based physical mapping (Soderlund et al. 2000, Nelson and Soderlund 2005). Comparison with a genetic map and the genomic scaffolds validates our physical map that comprises 3,312 BAC contigs and spans ~3,500 Mbp (97%) of the 3,600 Mbp sunflower genome.