Methods and reaction mixture reagent for increasing the...

Details Methods and reaction mixture reagent for increasing the... Methods and reaction mixture reagent for increasing the...

2007-12-25

2007-12-25

Kim, Young J. (Department: 1637)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

C435S006120

Reexamination Certificate

active

10867994

ABSTRACT:
A method for reducing the efficiency of primer extension by polymerase enzymes when the 3′ end of a primer or growing nucleic acid chain does not hybridize perfectly with the target, for increasing the selectivity of single nucleotide mutation or gene analyses, for suppressing false positive results and for enhancing the fidelity of the amplification of nucleic acid fragments by avoiding the incorporation of mispairs, the method comprising the steps of: (a) obtaining a nucleic acid sample; (b) hybridizing said nucleic acid sample to a primer; (c) subjecting said nucleic acid sample hybridized to a extension reaction by extending a primer with a polymerizing enzyme; and (d) detecting the presence of extension products; wherein the reaction extension mixture medium contains an intercalating agent such as ethidium bromide, dihydroethidium, ethidium homodimer-1, ethidium homodimer-2, acridine, propidium iodide, YOYO®-1 or TOTO®-1. When the intercalating agent is ethidium bromide the concentration is about 1 to 7 g/ml.

REFERENCES:
patent: 5976842 (1999-11-01), Wurst
patent: 6171785 (2001-01-01), Higuchi
patent: 0 512 334 (1992-11-01), None
patent: 1 256 631 (2002-11-01), None

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