Synthesis of error-minimized nucleic acid molecules
US7704690B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Dec 4, 2006 |
| Grant date | Apr 27, 2010 |
| Priority date | — |
| Expiry date | Dec 4, 2026 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1027
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method is provided for synthesis of error-minimized nucleic acid molecules. Oligonucleotides intended to have fragments of a desired, full-length nucleotide sequence, and optionally containing other desired nucleotides, such as nucleotides for binding the oligonucleotides to a substrate, are obtained. Oligonucleotides for both strands of the desired, full-length sequence may be obtained. The oligonucleotides are amplified and assembled into a first set of molecules intended to have the desired, full-length nucleotide sequence. The first set of molecules is denatured and annealed to form a second set of molecules intended to have the desired, full-length nucleotide sequence. The second set of molecules is cut into smaller segments, for example, by mixing the molecules with endonucleases that form blunt cuts in the second set of molecules where there are sequence errors, as well as randomly along the molecules. The smaller segments are assembled into a set of molecules intended to have the desired, full-length nucleotide sequence. By promoting cutting of the molecules in this manner near the end of the nucleic acid molecule synthesis process, a set of full-length molecules can be o…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.