Method of detecting sequence variations in macromolecules by thermal gradiant
AssigneeVarian, Inc.
InventorsReinhold E. Gerber, Richard G. Hatch
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Abstract
The present invention describes a method for separating macromolecules, particularly heteroduplex from homoduplex DNA molecules. The macromolecules undergo a temperature sensitive structural denaturation which, when it occurs, results in a difference in the mobilities of the denatured components on a solid support. A polar to non-polar solvent gradient or modified solid support particular to each sample is not required, since the difference in mobility is achieved on conventional solid supports using a single mobile phase composition.
Claims14
A chromatographic method for separating macromolecules in a mixture which undergo a temperature sensitive structural denaturation, comprising the steps of:
- applying a mixture containing said macromolecules onto a stationary reverse phase support under conditions which do not substantially denature said macromolecules;
- eluting said macromolecules with a mobile phase while exposing said macromolecules to a temperature gradient effective to at least partially denature said macromolecules, which results in the separation of said macromolecules.
A chromatographic method for separating macromolecules in a mixture which undergo a temperature sensitive structural denaturation, comprising the steps of:
- applying a mixture containing said macromolecules to a stationary reverse phase support under conditions which at least partially denature said macromolecules;
- eluting said macromolecules with a mobile phase while exposing said macromolecules to a temperature gradient, which results in the separation of said macromolecules.
Dependencies are inferred from the claim text.