TY - JOUR A1 - Zajac, Barbara Karolina A1 - Amendt, Jens A1 - Verhoff, Marcel A. A1 - Zehner, Richard T1 - Dating pupae of the blow fly calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for post mortem interval—estimation : validation of molecular age markers T2 - Genes N2 - Determining the age of juvenile blow flies is one of the key tasks of forensic entomology when providing evidence for the minimum post mortem interval. While the age determination of blow fly larvae is well established using morphological parameters, the current study focuses on molecular methods for estimating the age of blow flies during the metamorphosis in the pupal stage, which lasts about half the total juvenile development. It has already been demonstrated in several studies that the intraspecific variance in expression of so far used genes in blow flies is often too high to assign a certain expression level to a distinct age, leading to an inaccurate prediction. To overcome this problem, we previously identified new markers, which show a very sharp age dependent expression course during pupal development of the forensically-important blow fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) by analyzing massive parallel sequencing (MPS) generated transcriptome data. We initially designed and validated two quantitative polymerase chain reaction (qPCR) assays for each of 15 defined pupal ages representing a daily progress during the total pupal development if grown at 17 °C. We also investigated whether the performance of these assays is affected by the ambient temperature, when rearing pupae of C. vicina at three different constant temperatures—namely 17 °C, 20 °C and 25 °C. A temperature dependency of the performance could not be observed, except for one marker. Hence, for each of the defined development landmarks, we can present gene expression profiles of one to two markers defining the mentioned progress in development. KW - forensic entomology KW - digital age determination KW - qPCR KW - transcriptome analysis KW - metamorphosis KW - forensics KW - blow fly Y1 - 2018 UR - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/45866 UR - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-458668 SN - 2073-4425 N1 - This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0). VL - 9 IS - 3, Art. 153 SP - 1 EP - 19 PB - MDPI CY - Basel ER -