Use of immobilized mismatch binding protein for detection of mut

Details Use of immobilized mismatch binding protein for detection of mut Use of immobilized mismatch binding protein for detection of mut

1996-03-04

2000-09-19

Myers, Carla J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, C12Q 168, C12P 1934

Patent

active

061209929

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.

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