Use of immobilized mismatch binding protein for detection of mutations and polymorphisms, and allele identification in a diseased human
US6120992A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | Mar 4, 1996 |
| Grant date | Sep 19, 2000 |
| Priority date | — |
| Expiry date | Mar 4, 2016 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6827
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for detecting mutations, such as a single base change or an addition or deletion of about one to four base pairs, is based on the use of an immobilized DNA mismatch-binding protein, such as MutS, which binds to a nucleic acid hybrid having a single base mismatch or unpaired base or bases, thereby allowing the detection of mutations involving as little as one base change in a nucleotide sequence. Such a method is useful for diagnosing a variety of important disease states or susceptibilities, including the presence of a mutated oncogene and the presence of DNA containing triplet repeat sequences which characterize several genetic diseases including fragile X syndrome. The present method is used to isolate or remove by affinity chromatography duplex DNA molecules containing mismatches such as error-containing molecules in PCR-amplified DNA samples. Methods for detecting and enriching minority sequences are disclosed. Also provided are compositions and kits useful for practicing the methods of the present invention.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.