ABSTRACT

DNA has emerged as a promising storage medium for addressing exponentially growing data storage demands, owing to its exceptional information density and chemical stability. However, current DNA synthesis techniques face significant limitations in achieving high-throughput data storage due to constrained synthesis yields. This study presents a novel high-yield cap-free synthesis (HYCFS) strategy that overcomes the limitations of conventional solid-phase phosphoramidite chemistry in both synthesis length and production yield. We established a theoretical product prediction model based on cap-free synthesis characteristics and systematically evaluated the strategy through standard DNA storage workflows. Under column-based synthesis conditions with high coupling efficiency (>99%), this approach demonstrated a 3-fold enhancement in effective sequence yield compared to traditional methods. Theoretical modeling predicts superior performance in array-based synthesis systems for large-scale data storage applications. HYCFS shows potential to enhance DNA-based data storage capacity by 2 orders of magnitude while reducing storage costs, thereby advancing the development of large-scale DNA data storage technologies.