Patent ID: 11913071
Assignee: SINGULAR GENOMICS SYSTEMS, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 26:
27. A method of amplifying a circular polynucleotide, the method comprising:
(a) contacting a solid support with the circular polynucleotide and amplifying the circular template polynucleotide by extending a first primer attached to the solid support with a strand-displacing polymerase to generate a first immobilized extension product comprising one or more complements of the circular polynucleotide;
(b) contacting the first immobilized extension product with a second primer attached to the solid support and extending the second primer with a polymerase to generate a second immobilized extension product, wherein the second immobilized extension product is hybridized to the first immobilized extension product;
(c) hybridizing an invasion primer to the first immobilized extension product, wherein the invasion primer is not covalently attached to the solid support;
(d) extending the invasion primer hybridized to the first immobilized extension product with a polymerase to generate an invasion strand, thereby displacing the second immobilized extension product; and
(e) contacting the second immobilized extension product with a third primer and extending the third primer with a polymerase to generate immobilized amplification products.