Nucleic Acid Amplification via Immobilized Primers

The patent segments the amplification process by spatially separating multiple template-primer reactions onto different solid supports (beads, microparticles, or array positions). Each support unit contains or is associated with specific templates and primers, physically isolating them from other reactions. This segmentation enables multiplexed amplification of multiple targets simultaneously while preventing cross-contamination, as each reaction occurs in its own confined space on the support surface.

The patent employs self-service mechanisms where the solid support structure inherently provides the necessary functions for maintaining clonality. The immobilized primers on the support surface automatically ensure that amplification occurs only at the specific location where the primer is bound, without requiring additional external controls or intervention. The physical confinement of the reaction to the support surface and the co-localization of templates and primers on the same support unit naturally maintain clonal populations, eliminating the need for complex external control systems.

The patent utilizes parameter changes in the attachment process, specifically controlling the stringency conditions (temperature, salt concentration, pH) of hybridization and washing steps to optimize both template retention and cross-contamination prevention. By carefully adjusting these parameters, the system achieves strong specific binding of templates to their complementary primers on the support while allowing non-specific binding and cross-contaminants to be washed away. The immobilized primers on the support provide a stable anchor that prevents template loss during these parameter-adjusted washing steps.