Combinatorial Nanoparticle-Bound\nssDNA Oligonucleotide\nLibrary Synthesized by Split-and-Pool Synthesis

Synthetic ssDNA oligonucleotides hold great potential\nfor various\napplications, including DNA aptamers, DNA digital data storage, DNA\norigami, and synthetic genomes. In these contexts, precise control\nover the synthesis of the ssDNA strands is essential for generating\ncombinatorial sequences with user-defined parameters. Desired features\nfor creating synthetic DNA oligonucleotides include easy manipulation\nof DNA strands, effective detection of unique DNA sequences, and a\nstraightforward mechanism for strand elongation and termination. In\nthis study, we present a split-and-pool method for generating synthetic\nDNA oligonucleotides on nanoparticles, enabling the creation of scalable\ncombinatorial libraries. Our approach involves coupling DNA to nanoparticles,\nligating double-digested fragments for orientation-specific synthesis,\nand attaching a final single-digested fragment to ensure strand termination.\nWe assess the quality of our method by characterizing both the DNA\nand the nanoparticles used as solid supports, confirming that our\nmethod produces scalable, combinatorial nanoparticle-bound ssDNA libraries\nwith controllable strand lengths.