Data Storage Based on Combinatorial Synthesis of DNA Shortmers - Proof of concept
To assess and establish the potential of large-scale combinatorial alphabets, we combined simulations of large-scale combinatorial DNA systems and a smaller-scale experimental proof of concept. A Gibson assembly was used to construct two barcodes, each in four cycles that can cover 625 possible combinations, with a building block of twenty bases in length and an overlap of 20 bp between adjacent fragments. 16 building blocks are designed with two differing adjacent fragments that overlap. In four cycles, a total of 64 building blocks are constructed for every barcode, with a building block of twenty bases in length and an overlap of 20 bp between adjacent fragments. The purified assembly products are amplified using index primers for Illumina, relying on P5 and P7 anchors on 5' and 3' end, respectively. The size of the expected product is ~370bp