Abstract: Hemidactylus turcicus is a member of one of the most species-rich reptile genera and is known as an invasive species closely associated with humanized environments. Its phylogeography is often interpreted to be the result of recurrent human-mediated introductions. Çanakkale is the place that acts as a species corridor for the Anatolia peninsula. Farther, this place is connected with an adjacent island. Here, we tender the genetic structure of H. turcicus from both islands (Bozcaada) and the mainland of Çanakkale using mtDNA (Cyt b and COI gene regions). We used molecular tools to assess species identity and compare our sequences to a database containing various expertly identified species. We also applied DNA barcoding to report and release our data to the public. DNA barcoding is a reliable way to identify and store data on invasive species. Not surprisingly, specimens from Çanakkale genetically resemble other Mediterranean conspecifics from Italy, Greece, and Albania. Albeit, we found quite genetic differences between samples from Çanakkale and Adana. Not all cryptic lineages have been investigated taxonomically in H. turcicus. Researchers should implement genetic monitoring of specimens from its all-distributed area.