Work

Efficiency of Digital Photolithographic Synthesis of Large, High-Quality DNA Libraries and Microarrays using a Guanine O6 Dephosphitylation Strategy

Year: 2025

Type: article

Abstract: Large-scale de novo nucleic acid synthesis is a powerful tool enabling researchers to better understand and engineer biological systems. Fields ranging from genomics to nucleic acid therapeutics to synthetic biology make use of high-throughput experimental approaches requiring access to large pools or libraries of DNA, RNA, synthetic nucleic acid analogs, non-nucleosidic building blocks, or combinations of these. Large oligonucleotide libraries are synthesized as microarrays and used in situ for surface-based assays or cleaved for off-array applications. Here, using a digital maskless photolithographic approach, we address an important source of error in DNA microarray synthesis, oligonucleotide fragmentation arising from the O 6 -phosphitylation of guanine during the potentially hundreds of coupling cycles required for complex library synthesis. Introducing a very short debranching step using standard capping reagents suppresses depurination-based fragmentation and greatly enhances synthetic yield.

Source: Communications Chemistry

DOI: 10.1038/s42004-025-01755-0