Circularized Nucleic Acid Sequencing for Rare Variant Detection

The patent applies preliminary action by circularizing the nucleic acid template before sequencing and performing rolling circle amplification to generate multiple copies of the template. This pre-amplification step enriches the target sequence, enabling detection of rare variants with higher sensitivity while maintaining specificity through the structured amplification process that preserves sequence fidelity.

The patent uses an adapter molecule as an intermediary that facilitates the circularization of the nucleic acid template. The adapter contains specific sequences that enable binding and circularization, acting as a mediator between the linear template and the circularized form required for rolling circle amplification, thereby improving detection sensitivity without compromising specificity.

The patent extracts the amplification function into a separate rolling circle amplification step that occurs before sequencing. By isolating the amplification process from the sequencing process, the method achieves high sequencing accuracy through multiple template copies while managing complexity by separating the two functions into distinct operational phases.

The rolling circle amplification process generates a continuous growing strand that serves as the sequencing template. This continuous amplification produces multiple copies of the target sequence in a single ongoing reaction, improving sequencing accuracy through redundancy while maintaining a streamlined process that avoids multiple discrete steps.