Shotgun Sequencing of 512‐mer Copolyester Allows Random Access to Stored Information

Digital information encoded in polymers has been exclusively decoded by mass spectrometry. However, the size limit of analytes in mass spectrometry restricts the storage capacity per chain. In addition, sequential decoding hinders random access to the bits of interest without full‐chain sequencing. Here we report the shotgun sequencing of a 512‐mer sequence‐defined polymer whose molecular weight (57.3 kDa) far exceeds the analytical limit of mass spectrometry. A 4‐bit fragmentation code was implemented at aperiodic positions during the synthetic encoding of 512‐bit information without affecting storage capacity per chain. Upon activating the fragmentation code, the polymer chain splits into 18 oligomers, which could be individually decoded by tandem‐mass sequencing. These sequences were computationally reconstructed into a full sequence using an error‐detection method. The proposed sequencing method eliminates the storage limit of a single polymer chain and allows random access to the bits of interest without full‐chain sequencing. We demonstrated shotgun sequencing of 512 repeating unit sequence‐defined polymers with a molecular weight (57.3 kDa) far exceeding the analysis limit of mass spectrometry. Chemically activated fragmentation code was employed to generate MS‐compatible oligomers. Shotgun sequencing also allowed random access to the stored information.