Next Generation Sequencing workflows include a number of critical steps which can have a significant impact both on the quality of the results and on the overall cost of the project. In particular, DNA fragmentation and library preparation, as the first two steps of the workflow, are crucial, since at these early stages significant bias can be introduced to the sample, compromising the quality of the datasets obtained later. In this application note we describe how a fully functional, high-quality library for Illumina platforms has been prepared, using the Diagenode Bioruptor® Pico and the Bioline JetSeq™ Flex DNA Library Preparation Kit.