Comprehensive method development for a wide size range of RNA

The integrity and quality of the encapsulated nucleic acids used in cell and gene therapy are important to ensure the efficacy and safety of the therapeutic product. Current methods used to characterise the integrity and quality of these nucleic acids are complex and yield suboptimal analytical resolution and poor transferability. This work introduces a new chemistry and methods optimisation strategy that is applicable to single-stranded RNAs with lengths ranging from 50 to 9000 bases, enabling scientists to achieve high-resolution and high-quality data.

Introduction

This technical note showcases the reproducibility for electrokinetic injection, a %RSD for migration time below 0.25% and %RSD for corrected peak area% less than 3 for fragments between 300 and 2000 bases for the single-stranded RNA (ssRNA) ladder.

Compared to traditional slab gel-based electrophoresis methods, capillary gel electrophoresis (CGE), combined with laser-induced fluorescence (LIF), offers superior resolution, shorter analysis time, automated operation and exceptional sensitivity. Compared to chip-based CE systems, we demonstrate easy method modification and optimisation flexibility, allowing scientists to choose between project-specific methods to achieve optimal results and platform methods for higher throughput.

This study evaluated an RNA ladder that ranged from 50 to 9000 bases. The effects of varying conditions, such as injection mode and separation temperature, are described.

Key features

• Ready-to-use kit and method for single-stranded nucleic acid analysis
• Easy-to-optimise method strategy for samples containing small or large ssRNA fragments
• Ability to use either hydrodynamic or electrokinetic injections, depending on sample composition, and achieve great resolution
• Ability to separate a wide range of RNAs in a single separation
• Reproducible and reliable results from hydrodynamic injections, with migration time %CV <0.15% and corrected peak area% <4.5%
• Size estimation of RNA samples with easy-to-use and flexible modelling methods

Would you like to read more application notes?