DNA conjugation on functionalized plastic surfaces for sequential, iterative single molecule sequencing.

Area of Science:

• Biotechnology
• Materials Science
• Molecular Biology

Background:

• DNA offers high-density data storage potential.
• Current methods for DNA retrieval can be destructive or inefficient.
• Need for stable, reusable DNA data storage systems.

Purpose of the Study:

• To develop a method for repeated and sequential retrieval of arbitrary DNA elements.
• To create a robust, plastic-based DNA storage medium.
• To compare PCR and recombinase polymerase amplification (RPA) for DNA retrieval.

Main Methods:

• Conjugating synthetic DNA, tailed with trans-cyclooctene (TCO), to methyltetrazine (MTz)-functionalized plastic surfaces using click chemistry.
• Utilizing polymerase-based amplification (PCR and RPA) for DNA retrieval.
• Characterizing the DNA conjugation process, kinetics, and performing amplification and sequencing over multiple retrieval cycles.

Main Results:

• Demonstrated covalent DNA attachment to plastic surfaces enabling repeated, non-destructive amplification.
• Successfully retrieved and sequenced specific DNA data elements multiple times from the same surface.
• RPA exhibited better performance metrics, including reduced contamination and improved yield, compared to PCR.

Conclusions:

• Developed a plastic-based DNA storage system for robust, iterative, targeted DNA retrieval.
• The click chemistry approach allows for stable, long-term DNA data storage.
• RPA is a more suitable amplification method for sequential DNA data retrieval from this storage system.