Nucleic acid amplification
First Claim
1. A method for nucleic acid amplification, comprising:
- a) distributing at least two different polynucleotide templates onto an array of reaction sites on a support wherein the array of reaction sites comprise the same first universal primer that does not include a template sequence by introducing a single one of each of said different polynucleotide templates into at least two different reaction sites of the array, wherein the polynucleotide templates each include the same first universal primer binding site, the same second universal primer binding site, and are present within a reaction mixture in a single continuous liquid phase; and
b) forming at least two substantially monoclonal nucleic acid populations by amplifying, by partially denaturing and without compartmentalizing and within the same reaction mixture of step (a) in the continuous liquid phase, each of the single polynucleotides within said at least two reaction sites, wherein the amplifying is performed using a second universal primer in solution that can hybridize to the second universal primer binding site within the polynucleotide templates, and wherein said at least two reaction sites are in fluid communication with each other during the amplifying.
2 Assignments
0 Petitions
Accused Products
Abstract
In some embodiments, the present teachings provide methods for nucleic acid amplification, comprising forming a reaction mixture, and subjecting the reaction mixture to conditions suitable for nucleic acid amplification. In some embodiments, methods for nucleic acid amplification include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, methods for nucleic acid amplification include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the methods for nucleic acid amplification employ an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel. In some embodiments, methods for nucleic acid amplification can be conducted in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components. In some embodiments, methods for nucleic acid amplification comprise a amplifying at least one polynucleotide onto a surface under isothermal amplification conditions, optionally in the presence of a polymer. The polymer can include a sieving agent and/or a diffusion-reducing agent.
-
Citations
20 Claims
-
1. A method for nucleic acid amplification, comprising:
-
a) distributing at least two different polynucleotide templates onto an array of reaction sites on a support wherein the array of reaction sites comprise the same first universal primer that does not include a template sequence by introducing a single one of each of said different polynucleotide templates into at least two different reaction sites of the array, wherein the polynucleotide templates each include the same first universal primer binding site, the same second universal primer binding site, and are present within a reaction mixture in a single continuous liquid phase; and b) forming at least two substantially monoclonal nucleic acid populations by amplifying, by partially denaturing and without compartmentalizing and within the same reaction mixture of step (a) in the continuous liquid phase, each of the single polynucleotides within said at least two reaction sites, wherein the amplifying is performed using a second universal primer in solution that can hybridize to the second universal primer binding site within the polynucleotide templates, and wherein said at least two reaction sites are in fluid communication with each other during the amplifying. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
-
Specification