Enzyme-based regeneration of surface-attached nucleic acids
US7101669B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 12, 2001 |
| Grant date | Sep 5, 2006 |
| Priority date | — |
| Expiry date | Apr 4, 2022 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6837
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Enzyme-based regeneration of surface-attached nucleic acids is described herein. Microarrays involving hybridization of a probe to a target are important tools for genetic analysis. Conventionally, a microarray is used for a single analysis, after which it is discarded. The invention relates to a process for regeneration of a microarray through enzymatic digestion of a target from a surface-attached probe using a nuclease to digest a single strand of a nucleic acid duplex with directional specificity starting from the free end of the target strand. For example, a probe oligonucleotide bound to a gene chip at the 5′-end hybridizes to a target nucleic acid, leaving the 5′ end of the target open to 5′–3′ digestion. Lambda-exonuclease (λ-exonuclease) cleaves single nucleotides from the 5′ end of a duplex, progressing in the 5′–3′ direction. Once the target strand is digested, the enzyme is rinsed from the microarray. The microarray is thus regenerated and ready for a subsequent use.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.