This study investigates the effect of tarantula species on invasive species using Cytochrome c oxidase I (COI) gene. DNA was extracted from tarantula samples provided by the Kansas State University’s Insect Zoo, Disney’s Animal Kingdom, and other institutions. The DNA was analyzed through PCR amplification, gel electrophoresis, and DNA sequencing using the DNA Subway and NCBI BLAST platforms. By identifying and understanding species diversity through DNA barcoding, researchers can better protect endangered or ecologically important organisms and help maintain balanced ecosystems.