Quality Control of Chemically Synthesized sgRNA Using ONT DRS

Existing quality control methods for RNA synthesis can only provide insight towards RNA length distribution or limited composition information for short oligos. We developed a standard quality control metric using Oxford Nanopore direct RNA sequencing to obtain direct insight into RNA length distribution, sequence, and presence of RNA modification sites. Our pipeline can identify error-prone regions and truncation sites that occur during synthesis. Furthermore, problematic steps in the synthesis are identified and repaired. We show that our platform can produce and assess 101-mer single guide RNA with high-fidelity and show that modifications can be reliably detected.