The biological clock can influence the behavior and physiology of organisms. In exploring differences in circadian rhythm characteristics, past studies often focused on strains from North America, lacking an investigation into the circadian rhythm characteristics of strains from the equator and the Southern Hemisphere. The Drosophila melanogaster is a widely distributed specie across various continents. Thus, we used Drosophila (equatorial and mid-high latitude strains) as the subject to study whether flies exhibit different circadian rhythm characteristics due to latitude. The results show that Drosophila strains from different latitudes exhibit distinct circadian rhythm characteristics. Equatorial strains demonstrate a more stable rhythm in complete darkness like the one in LD 12:12, while mid- high latitude strains from the Northern and Southern Hemispheres share similar rhythm characteristics, showing an unstable rhythm that does not align with the stable rhythm under normal light conditions in complete darkness. Additionally, there are strain-specific differences in activity levels, period, and rhythm strength. Further comparison of gene sequences in various strains in the experiment revealed many SNP variations in the regulatory regions of per and tim, indicating a relationship with circadian rhythm characteristics, in order to identify potential genetic variations responsible for the differences in circadian rhythm characteristics.