The introduction of DNA barcoding has highlighted the expanding use of the mtDNA gene, cytochrome c oxidase I (COI), as a genetic marker for species identification (Dawnay et al., 2007). In the current study, COI gene based barcoding was employed to identify the seven spotted cockroach, Therea petiveriana. The genetic diversity amongst this cockroach collected from various locations in Tamil nadu, India was analysed by the banding patterns obtained in RAPD PCR. This was further validated using PCR RFLP studies on the mtDNA gene, COI. The results highlighted the impact of varying environmental factors on the genetic makeup of these species.