Patterned Oligonucleotide Arrays via Enzymatic Transfer

The patent uses template arrays with oligonucleotide sequences that serve as templates for synthesizing complementary probes on the recipient array. The template arrays are copied onto the recipient array through hybridization and extension reactions, producing high-quality full-length probes. This copying approach ensures manufacturing precision while maintaining cost-effectiveness through reusable templates.

The patent employs pre-synthesized template arrays that are prepared in advance with known sequences and orientations. These templates are then used to guide the synthesis of complementary probes on the recipient array. The preliminary preparation of templates with correct orientations and sequences ensures that the final probes achieve high manufacturing precision without requiring complex real-time synthesis procedures.

The patent implements directional control at the local level by designing template oligonucleotides with specific orientations (5′ to 3′ directionality) that ensure complementary probes are synthesized in the correct orientation for enzymatic reactions. Each probe on the recipient array is oriented uniformly, with the 5′ end attached to the surface and the 3′ end extending outward, optimizing local enzymatic reaction efficiency without requiring complex global reconfiguration of the fabrication process.

The patent utilizes asymmetric orientation of oligonucleotide probes on the array surface. The probes are synthesized with a specific asymmetric configuration where the 5′ end is anchored to the recipient array surface while the 3′ end remains free in solution. This asymmetric arrangement is crucial for enzymatic reactions such as extension and sequencing, as it allows enzymes to access the 3′ end efficiently. The asymmetric design is achieved through the template-directed synthesis process, which inherently produces probes in a uniform orientation.