Differential gene expression for age estimation of forensically important Sarcophaga peregrina intra-puparial (Diptera: Sarcophagidae)

   Sarcophaga peregrina (Diptera: Sarcophagidae) is an important flesh fly species for estimation the minimum postmortem interval (PMImin) in forensic entomology. The accurate determination of the developmental age is a crucial task for using necrophagous sarcophagids to estimate PMImin. During larval development, the age determination is straight forward by the morphological changes and variation of length, weight, and width, however, the age estimation of sarcophagid Intra-puparial is more difficult due to anatomical and morphological changes not being visible. The analysis of differentially expressed genes (DEGs) during sarcophagid metamorphosis is a potential method for age estimation of Intra-puparial. In the present study, real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the differential gene expression level of S. peregrina Intra-puparial in different constant temperatures (35°C, 25°C and 15°C). In addition, the appropriate reference genes of S. peregrina were selected in the Intra-puparial and at different temperatures to obtain reliable and valid gene expression profiles.