An electric field and runtime driven band model for high-speed, real-time imaging gel electrophoresis

Background: Traditional temperature-dependent models face difficulties in compact systems due to complex temperature control. This study introduces an electric field strength and runtime driven (E-t) band model to improve GE performance by correlating band behavior with electric field and runtime rather than temperature. Results: We developed a compact E-t model based GE system, which equipped with inert platinum-titanium electrodes, a quartz-glass-embedded tank for passive cooling, and smartphone-based real-time fluorescence imaging. Experimental results from real-time tracking of GE bands, model fitting under different E-t conditions, and the separation of rice receptor protein kinase genes (CERK1 and CEBiP) confirmed that the proposed model can accurately describe electrophoretic band migration and dispersion, while maintaining good agreement with traditional temperature-based models and being little affected by temperature. Furthermore, successful nucleic acid separation was achieved within minutes under a high electric field strength in our system. Significance: By minimizing the reliance on temperature control mechanisms, the E-t band model offers a new perspective for the design of analytical chemistry instruments, enabling electrophoresis to focus primarily on optimizing the two key parameters, E and t. In addition, our portable, real-time imaging GE system enhances separation efficiency and provides a practical, high-performance reference solution for rapid, on-site analysis applications.