Background: The precise identification of forensic insects is one of the most important aspects in forensic entomology since it is the basis for determining post-mortem intervals (PMI) and providing other valuable forensic evidence in criminal investigations. Methods: This study demonstrates the utility of DNA barcoding in accurately identifying new strains of forensic insects collected from the Jeddah Governorate. Result: High similarity percentages ranging from 98.74% to 99.68% were observed, underscoring the precision of molecular identification. The identified species included members of the Diptera (flies) and Coleoptera (beetles) orders, commonly associated with forensic cases. Notably, Phoridae sp. (Forensic_JED_21) was identified with 98.74% similarity, highlighting their role in providing valuable post-mortem interval (PMI) estimations. Physiphora alceae was identified twice (Forensic_JED_22 and Forensic_JED_24) with similarities of 99.22% and 99.07%, respectively, demonstrating the robustness of DNA barcoding. Philonthus discoideus (Forensic_JED_23) and Physiphora demandata (Forensic_JED_25) were identified with 99.68% similarity, confirming the presence of these species in the Jeddah region. Phylogenetic analysis using MEGA11 placed these strains in well-defined clades, confirming their taxonomic accuracy and evolutionary relationships. These findings align with previous research on the genetic diversity of forensic insects, adding crucial data to forensic entomology databases. This study enhances the forensic entomology framework by providing accurate species identification, essential for legal investigationsand supports the development of region-specific forensic tools through the addition of new genetic data from Jeddah.
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