The identification of immature Diptera is important in forensic entomology because it allows an estimation of the time and location of death. Previously, identification of Diptera larvae has relied on variations in spiracle patterns and the cephalopharyngeal skeleton. This type of analysis requires an expert in the field to assure a correct identification. In this study, the authors tested a simple, rapid method that utilized RFLP’s within the internal transcribed spacer (ITS) regions of the ribosomal DNA gene to identify 10 forensically important dipteran species in three families (Calliphoridae, Muscidae, and Sarcophagidae). The region from 18S to the 28S rRNA genes was amplified using polymerase chain reaction (PCR). Findings were that the ITS1 and ITS2 regions showed variation between species and homogeneity within species, with the exception of only one species. Identification of the three families of Diptera was provided by combinations of the restriction enzymes DdeI, Hinf1, and Sau3AI. Due to regional variation, future research should focus on determining RFLP patterns for additional dipteran species. Reference