Purifying Nucleic Acids through Redox-Responsive Polyacrylamide Gel Electrophoresis with Enhanced Recovery

Polyacrylamide gel electrophoresis (PAGE) is widely used for the analysis and purification of macromolecules, including nucleic acids and proteins. However, product recovery from conventional PAGE remains inefficient due to the densely cross-linked gel matrix. In this study, we develop a redox-responsive polyacrylamide gel by partially substituting the traditional cross-linker with a redox-cleavable alternative. The resulting gel retains separation performance comparable to standard PAGE, while exhibiting pronounced redox-triggered swelling, which facilitates the efficient release of nucleic acids under reducing conditions. Based on this strategy, DNA recovery efficiencies were increased up to ∼63%, more than 2-fold higher than traditional PAGE, with minimal residual polymer contamination. The platform also demonstrated applicability for RNA and proteins with increased recovery. These results suggest that this redox-responsive system will provide a complementary approach for macromolecular collection in chemical and biological research.