A database for simulation of double digest selective label (DDSL) typing technique has been created and validated against a sequenced strain (Salmonella enterica serovar Typhimurium strain LT2). In silico bands were in agreement with experimental, and the technique was able to discriminate among strains belonging to the same species. When compared with other strain discrimination techniques, DDSL showed a higher discriminatory power. The database contains precomputed data which may be searched to retrieve experimental conditions for typing all up-to-dated sequenced prokaryotic microorganisms.