HONOURs Training “Direct Next Generation Sequencing (NGS) after shearing” at FLI

Dirk Höper, Patrick Zitzow and Pauline Santos welcomed five ESRs from the HONOURs partner institutes on June 18 (Monday) for a one-week-training in “Direct NGS after shearing” at the Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health at the headquarters located at the Baltic Isle of Riems. After the obligatory biosafety instructions, the trainees got an overview of the Direct NGS workflow and its theoretical background, followed by the practical course led by Patrick Zitzow, technician at the FLI NGS-Laboratory, and assisted by Pauline Santos. In this session, the trainees processed diagnostic samples, which were sent to the FLI NGS-Laboratory for diagnostic metagenomics analyses. Divided in two groups, the ESRs handled the samples according to the established FLI workflow, starting with sample disintegration, followed by RNA extraction, cDNA synthesis, shearing and then library preparation. Finally, they performed quality control of the size-selected libraries before they prepared the sequencing run. During the sequencing run, the ESRs participated on a seminar dealing with the detailed FLI metagenomics data analysis workflow (RIEMS). On the final day, they had the opportunity to follow the data processing of RIEMS in real time, and assessed the entire workshop in parallel. The highly motivated ESRs demonstrated excellent laboratory performance, wherein they finished the assigned tasks ahead of schedule. Their interest in the workflow design and understanding of the rationale behind the combination of the different steps were very good. Furthermore, they used all opportunities and the dedicated question and answer sessions for intense discussions.