Abstract

Eurofins Genomics Europe Pharma and Diagnostic Products & Services Synthesis GmbH is one of the world-leading oligonucleotide manufacturers for the industrial and research sector and a major provider for genotyping and DNA sequencing services. Several different classes of unmodified and modified oligonucleotides are produced via solid phase oligonucleotide synthesis in a high throughput production floor with short turnaround times (TAT). The portfolio ranges from custom DNAs and RNAs over fluorescent probes for qPCR applications to unique dual index (UDI) primer sets for state-of-the-art next generation sequencing (NGS) methods. Over the last decades, oligonucleotides have gained an important role in the field of biochemical research, diagnostics and therapeutic applications. Great effort has been made to optimize and automate oligonucleotide synthesis, especially to minimize undesired by-products such as truncated sequences resulting from the large number of required individual reactions. Since 2021, Eurofins Genomics can capitalize from next generation high throughput synthesizers, the so-called continuous flow synthesizers (CFS), which allow a simultaneous production of 6 x 384 different oligonucleotides with elevated speed reaching an excellent coupling efficiency of ≥99.6%. Here, we present quality control data of an unmodified crude poly-T 200mer with remarkable quality. Furthermore, single internally quenched (SIQ) qPCR probes with improved properties are presented. Based on the extensive knowledge Eurofins gained during the pandemic, these hydrolysis probes have been designed and their quality evaluated by using several analytical methods, such as UPLC and MS. The functionality has been examined in a SARS-CoV-2 model system and the resulting data show an impressive performance of the newly developed qPCR probes.