Development of workflows for metabarcoding of mass-samples

Abstract

The main goal was developing a workflow for comprehensive biodiversity monitoring using a DNA Barcoding approach. As one of the major insect groups in the Palearctic the diverse order of Diptera served as an experimental organism. A fundamental question was the suitability of cytochrome c oxidase (COI) as a entity marker. This included several steps: the collection of DNA data from previously identified species for the purpose of providing species references, testing the effectiveness of the marker, providing and evaluating appropriate mixed samples, working with MOTUs for a species independent evaluation and developing an automated trap. For cost-effectiveness and high throughput hybrid enrichment and methods of parallel sequencing were used to ensure high-quality data. An insight into the local species composition was obtained. From this the ecology and difference in diversity of two experimentally sampled forest types were derived. The results of this observation served as an exemplary demonstration of the workflow an a proof of concept.