Method of nucleic acid amplification

摘要

A nucleic acid molecule can be annealed to an appropriate immobilized primer. The primer can then be extended and the molecule and the primer can be separated from one another. The extended primer can then be annealed to another immobilized primer and the other primer can be extended. Both extended primers can then be separated from one another and can be used to provide further extended primers. The process can be repeated to provide amplified, immobilized nucleic acid molecules. These can be used for many different purposes, including sequencing, screening, diagnosis, in situ nucleic acid synthesis, monitoring gene expression, nucleic acid fingerprinting, etc.

主权项

1. A method for amplifying a nucleic acid derived from a sample derived from an individual on a solid support, comprising:
providing the nucleic acid; amplifying the nucleic acid with at least two primers by adding additional sequences to the nucleic acid thereby forming a modified nucleic acid, the at least two primers comprising the first primer comprising a first sequence portion identical to or complementary to a sequence of a third primer, a second sequence portion comprising a nucleic acid tag that identifies the origin of the nucleic acid, and a third sequence portion identical to or complementary to a portion of the nucleic acid; the second primer comprising a fourth sequence portion identical to or complementary to a sequence of a fourth primer and a fifth sequence portion identical to or complementary to a portion of the nucleic acid; amplifying the modified nucleic acid using the third and fourth primers, at least one of the third and fourth primers attached to the solid support, thereby forming a nucleic acid colony comprising extended third and fourth primers, and further amplifying the nucleic acids in the nucleic acid colony; wherein the fifth sequence portion of the second primer is configured to complement an extension product of the first primer on the nucleic acid, and the third sequence portion is configured to complement an extension product of the second primer on the nucleic acid.