An AmpliSeq™ HD approach for specific and sensitive identification of GMO by Next Generation Sequencing

Abstract

With new genetically modified crops being constantly developed, many new targets need to be analysed for regulatory compliance. Classical PCR-based approaches currently used are however reaching their limits in terms of target throughput and new approaches are needed to adapt to the evolving situation. The Ion AmpliSeq™ HD technology was here adapted for the simultaneous detection of 108 GMO targets by Next Generation Sequencing (NGS) on the Ion GeneStudio S5 platform, using PCR primers mostly retrieved from the official European GMO methods. Tested on a wide array of reference materials, the method delivered specific detection of the expected GMO targets and reached a sensitivity down to 0.01 % (w/w) as required by GMO analytical guidelines. Numerous proficiency test samples were successfully analysed which confirmed the method performance and its applicability on real test samples. With its straightforward library preparation and ready-to-use bioinformatics analysis, the developed method is well tailored for routine GMO testing laboratories.