We have developed an in silico model for simplified detection of five major food borne bacteria using common PCR primer based on conserved region °anking a variable region in the first rRNA operon sequences. The RFLP pattern exhibited on virtual SmaI and MboI digestion were unique to each genus. This design is based on the combination of comparative sequence analysis, conventional PCR and restriction digestion methods. We believe this approach as a better alternative to 16S rRNA sequencing based identification / detection of bacteria and suited for the resource limited academic research laboratories in which variety of bacteria are used in different students’ projects. © 2014, International Association of Scientists in the Interdisciplinary Areas and Springer-Verlag Berlin Heidelberg.