Repair of nucleic acids for improved amplification
US7700283B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 20, 2005 |
| Grant date | Apr 20, 2010 |
| Priority date | — |
| Expiry date | May 22, 2026 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6848
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI.The reaction mixture may further contain an AP endonuclease and a polymerase. These enzymes are optionally selected according to their ability to withstand high temperatures so they can be included in an amplification mixture. The reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.