Detection of mutations in nucleic acids by chemical cleavage

Details Detection of mutations in nucleic acids by chemical cleavage Detection of mutations in nucleic acids by chemical cleavage

1998-08-06

1999-10-26

Jones, W. Gary.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 536 231, 536 243, C12Q 168, C12P 1934, C07H 2102, C07H 2104

Patent

active

059726180

ABSTRACT:
Improved methods, compositions and kits for detecting mutations in nucleic acids are disclosed, such comprise annealing a piece of control nucleic acid without mutations to a piece of test nucleic acid very similar in sequence to the control nucleic acid but possibly containing mutations, treating this mixture with potassium permanganate or hydroxylamine to remove mismatched bases from the duplex nucleic acid, treating the resulting nucleic acid with any of a class of diamines, triamines, and tetraamines analogous to 1,2-ethylenediamine to cleave abasic sites, and then analyzing the chemically treated nucleic acid to determine whether cleavage has occurred and approximately at what position in the nucleic acid any cleavage has occurred. Inclusion of betaine in the base-removal reactions improves their sensitivity and specificity. This series of non-hazardous, non-noxious, chemical treatments can be performed rapidly in a single reaction tube with no or minimal intervening separation steps; and the cleavage product can be prepared for the analysis step without removing the chemical reagents used to effect cleavage.

REFERENCES:
patent: 4818268 (1989-04-01), Holtz
patent: 5217863 (1993-06-01), Cotton et al.

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