Development Of Purity Analysis Methods For Synthetic pegRNAs For Prime Editing Research Applications

Tingting Li, Sahana Mollah, Elliott Jones, Ashley Jacobi, Morgan Sturgeon, and Garrett Rettig, SCIEX, USA Integrated DNA Technologies, USA

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Achieve high editing efficiency and precision with Prime editing (PE). Here we outline a method that uses capillary gel electrophoresis to mitigate analytical quality control challenges for purity analysis, providing a quality assessment of chemically synthesized pegRNA within 30 minutes per sample. Develop a robust denaturing technique for pegRNA with high levels of secondary structure and achieve a fast, easy workflow for the purity analysis of complex pegRNA molecules.

Key features of pegRNA purity analysis include-

• Innovative purity analysis workflow for chemically synthesized pegRNAs with high-level secondary structures.

• Purity determination of pegRNA from the high-resolution separation of the full-length product from its impurities.

• A high level of repeatability with relative standard deviation% (RSD%) of <3% was achieved as a result of accurate and stable capillary temperature control on the PA 800 Plus system.
• Fast quality assessment of chemically synthesized pegRNAs within 30 minutes per sample