Ligation assembly and detection of polynucleotides on solid-support
First Claim
Patent Images
1. A method of synthesizing a polynucleotide on a solid-support, the method comprising the steps of:
- a. annealing one or more bridging oligonucleotides and two or more assembly oligonucleotides such that a ligatable nick site is formed between adjacent assembly oligonucleotides, wherein one of the assembly oligonucleotides is immobilized to a solid support;
b. ligating the ligatable nick sites thereby forming an immobilized ligation product;
c. annealing a primer to the immobilized ligation product; and
d. extending the primer to create an immobilized, double-stranded polynucleotide.
8 Assignments
0 Petitions
Accused Products
Abstract
The present invention concerns methods of assembly of a polynucleotide on a solid-support by performing steps of annealing, ligation, and extension. The steps may be repeated in a cyclical manner to assemble immobilized double- or single-stranded polynucleotides with functional gene properties. The immobilized polynucleotides may be amplified by the polymerase chain reaction, and detected and quantitated by an exonuclease assay with a self-quenching, fluorescent probe. The polynucleotide may be cleaved from the solid-support by chemical or enzymatic cleavage.
-
Citations
22 Claims
-
1. A method of synthesizing a polynucleotide on a solid-support, the method comprising the steps of:
-
a. annealing one or more bridging oligonucleotides and two or more assembly oligonucleotides such that a ligatable nick site is formed between adjacent assembly oligonucleotides, wherein one of the assembly oligonucleotides is immobilized to a solid support; b. ligating the ligatable nick sites thereby forming an immobilized ligation product; c. annealing a primer to the immobilized ligation product; and d. extending the primer to create an immobilized, double-stranded polynucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
-
Specification