Custom DNA Origami Scaffolds for Digital Nucleic Acid Memory

Motivated by the continuous growth of data storage capacity the semiconductor memory materials are approaching their production limits. DNA has been selected as a valid alternative candidate due to its information density and long retention time. Here at the Nucleic Acid Memory Institute we developed digital Nucleic Acid Memory (dNAM), which uses DNA origami as a breadboard for data storage that provides a matrix of complementary strands that bind dye-labeled single stranded DNA (ssDNA). DNA origami is made from a ssDNA scaffold and short oligonucleotides (short ssDNA) that fold together into a predefined shape. The dNAM project currently relies on the M13 ssDNA scaffold which has a fixed length of 7.2 kilobase pairs (kb), limiting size and customization of the DNA origami. Ability to create larger scaffolds will expand the storage capacity and versatility of the DNA origami breadboard. In order to create larger scaffolds, we used a phagemid/helper phage system to produce ssDNA in bacteria and then export it from the cell. Using restriction enzymes cloning we produced a custom 11,054 base pair scaffold from previous ssDNA produced phagemids. Future project’s perspectives are to increase yield of the scaffold production as long as its length and create a fully customizable sequence tool to expand the robustness and accuracy of dNAM