Express barcoding with NextGenPCR and MinION for species-level sorting of ecological samples.

The use of DNA barcoding is well established for specimen identification and large-scale biodiversity discovery, but remains underutilized for time-sensitive applications such as rapid species discovery in field stations, identifying pests, citizen science projects, and authenticating food. The main reason is that existing express barcoding workflows are either too expensive or can only be used in very well-equipped laboratories by highly-trained staff. We here show an alternative workflow combining rapid DNA extraction with HotSHOT, amplicon production with NextGenPCR thermocyclers, and sequencing with low-cost MinION sequencers. We demonstrate the power of the approach by generating 250 barcodes for 285 specimens within 6 h including specimen identification through BLAST. The workflow required only the following major equipment that easily fits onto a lab bench: Thermocycler, NextGenPCR, microplate sealer, Qubit, and MinION. Based on our results, we argue that simplified barcoding workflows for species-level sorting are now faster, more accurate, and sufficiently cost-effective to replace traditional morpho-species sorting in many projects.

Mission possible: an optimised protocol for the unbarcodable Ceraphronoidea (Hymenoptera).

DNA barcodes provide a reliable and efficient solution to resolving cryptic species complexes and accelerate species discoveries. The superfamily Ceraphronoidea (Hymenoptera) is a group of parasitoid wasps for which a barcoding approach could be of great help, if it were not for the very poor results. The inability to obtain barcodes for the majority of treated ceraphronoids halts progress on the taxonomy of this hyperdiverse parasitoid group. We here present a working protocol for the barcoding of ceraphronoid wasps which yields a first-time over 90% success rate.

New records of German Scelionidae (Hymenoptera: Platygastroidea) from the collection of the State Museum of Natural History Stuttgart.

BACKGROUND: Scelionid wasps are arthropod egg parasitoids, many of which are relevant to global biosecurity. However, the scelionid fauna of Germany has not received much attention from professional taxonomists. NEW INFORMATION: Eleven species and four genera are recorded for the first time from Germany, including species of interest to agriculture and biological control. First genus records include Baryconus Förster, Macroteleia Westwood, Paratelenomus Dodd and Probaryconus Kieffer. First species records include B.europaeus (Kieffer), Idrisnigroclavatus (Kieffer), Idrissemiflavus (Kieffer), M.bicolora Kieffer, M.pannonica Szabo, Paratelenomussaccharalis (Dodd), Trimorusvaricornis (Walker), Trissolcusbasalis (Wollaston), Trissolcusbelenus (Walker), Trissolcuscolemani (Crawford) and Trissolcusflavipes (Thompson). COI barcodes are identified for the first time from B.europaeus and M.bicolora. Each species is illustrated and updated world distributions are provided. Implications for agriculture are discussed.