| Summary: | <i>Sarcophaga peregrina</i> (Robineau-Desvoidy, 1830) has the potential to estimate the minimum postmortem interval (PMI<sub>min</sub>). Development data and intra-puparial age estimation are significant for PMI<sub>min</sub> estimation. Previous research has focused on constant temperatures, although fluctuating temperatures are a more real scenario at a crime scene. The current study examined the growth patterns of <i>S. peregrina</i> under constant (25.75 °C) and fluctuating temperatures (18–36 °C; 22–30 °C). Furthermore, differentially expressed genes, attenuated total reflectance Fourier-transform infrared spectroscopy, and cuticular hydrocarbons of <i>S. peregrina</i> during the intra-puparial period were used to estimate age. The results indicated that <i>S. peregrina</i> at fluctuating temperatures took longer to develop and had a lower pupariation rate, eclosion rate, and pupal weight than the group at constant temperatures did. Moreover, we found that six DEG expression profiles and ATR-FTIR technology, CHCs detection methods, and chemometrics can potentially estimate the intra-puparial age of <i>S. peregrina</i> at both constant and fluctuating temperatures. The findings of the study support the use of <i>S. peregrina</i> for PMI<sub>min</sub> estimation and encourage the use of entomological evidence in forensic practice.
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