DNA Barcoding in Forensic Entomology – Establishing a DNA Reference Library of Potentially Forensic Relevant Arthropod Species,

Abstract

Throughout the years, DNA barcoding has gained in importance in forensic entomology as it leads to fast and reliable species determination. High-quality results, however, can only be achieved with a comprehensive DNA barcode reference database at hand. In collaboration with the Bavarian State Criminal Police Office, we have initiated at the Bavarian State Collection of Zoology the establishment of a reference library containing arthropods of potential forensic relevance to be used for DNA barcoding applications. CO1-5P' DNA barcode sequences of hundreds of arthropods were obtained via DNA extraction, PCR and Sanger Sequencing, leading to the establishment of a database containing 502 high-quality sequences which provide coverage for 88 arthropod species. Furthermore, we demonstrate an application example of this library using it as a backbone to a high throughput sequencing analysis of arthropod bulk samples collected from human corpses, which enabled the identification of 31 different arthropod Barcode Index Numbers.

A DNA barcode library for 5,200 German flies and midges (Insecta: Diptera) and its implications for metabarcoding‐based biomonitoring

“…Previous studies have shown the great potential of metabarcoding for biotic assessments in various contexts, including Malaise trap surveys (Morinière et al, ), biosurveillance of invasive and pest species (Ashfaq & Hebert, ; L. A. Hardulak et al in prep), macrozoobenthos sampling for assessing water and stream health (Elbrecht & Leese, ; Serrana, Miyake, Gamboa, & Watanabe, ), faeces analyses for dietary inference (De Barba et al, ; Hawlitschek, Fernández‐González, Balmori‐de la Puente, & Castresana, ), species identification for forensic entomology (Chimeno et al, ) and for soil biology (Oliverio, Gan, Wickings, & Fierer, ). This approach combines the advantages of DNA barcoding, namely the capacity to identify any life stage, body fragment or even trace DNA in the environment, with the ability of high‐throughput sequencers to analyse millions of DNA fragments and thousands of specimens at a time.…”

“…Variation in overall biodiversity between sampling sites as well as annual phenologies of certain taxa can easily be visualized using presence-absence maps (Figure 2). This will be a useful feature for comparison of large data sets and for monitoring benefi- (Chimeno et al, 2018) and…”

Applications of forensic entomology: overview and update

“…Umożliwia ona analizę wielu osobników i generowanie dużych zbiorów danych. Choć nie została jeszcze w pełni przetestowana, w przyszłości może stać się ważnym narzędziem w badaniach na szerszą skalę [68,69].…”

“…The application of HTS enables the analysis of huge numbers of individuals as a whole and the generation of large datasets. Although this method was only tested on a small-scale, it is clear that HTS is an important tool for future large scale purposes [68,69].…”

The Challenge of DNA Barcoding Saproxylic Beetles in Natural History Collections—Exploring the Potential of Parallel Multiplex Sequencing With Illumina MiSeq

“…This started by a purification step of 400 µL of each pool of amplicons using NucleoMag 0.85X, then eluted in 50 µL TET buffer (0.1X), and followed by DNA quantification using Qubit R Broad Range. In contrast to Meyer and Kircher (2010), we performed a blunt-end repair using NEBNext End Repair Module before proceeding with a ligation step to attach Illumina adapters to our libraries and thus avoid an additional PCR step that may increase replication errors, especially when using a non-proofreading high-fidelity polymerase enzyme (Meyer et al, 2012;Leray et al, 2016;Chimeno et al, 2018). Approximately 500 ng of DNA were used with 5 µL of NEBNext Repair Reaction Buffer (10X) and 2.5 µL of NEBNext End Repair Enzyme Mix.…”