Chromatographic method for mutation detection using mutation site specifically acting enzymes and chemicals
First Claim
1. A method for analyzing a sample of double stranded DNA to determine the presence of a mutation site therein comprising:
- a) contacting said sample with a mutation site binding reagent which binds within the vicinity of said mutation site, andb) chromatographically separating and detecting the product of step (a) while the mutation site binding reagent remains bound to the mutation site.
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Abstract
A method for analyzing a sample of double stranded DNA to determine the presence of a mutation therein comprises contacting the sample with a mutation site binding reagent, and chromatographically separating and detecting the product. The chromatographic separation can be performed using Matched Ion Polynucleotide Chromatography, size exclusion chromatography, ion exchange chromatography, or reverse phase chromatography. The mutation site binding reagent can be an enzyme or a non-proteinaceous chemical reagent. In one embodiment, a mutation site binding reagent binds to the site of mutation and alters the chromatographic retention time. In another embodiment, a mutation site binding reagent cleaves at the site of mutation, resulting in an increase in the number of fragments.
67 Citations
31 Claims
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1. A method for analyzing a sample of double stranded DNA to determine the presence of a mutation site therein comprising:
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a) contacting said sample with a mutation site binding reagent which binds within the vicinity of said mutation site, and b) chromatographically separating and detecting the product of step (a) while the mutation site binding reagent remains bound to the mutation site. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A chromatographic method for analyzing a DNA sample to determine the presence of mutations in said sample, the method comprising:
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(a) separating said sample using a chromatographic method which produces a first chromatogram comprising peaks or other shapes which represent separated components of the sample; (b) contacting said sample with a mutation site binding reagent which binds within the vicinity of said mutation site; (c) separating the product of step (b) by the chromatographic method of step (a) to produce a second chromatogram while the mutation site binding reagent remains bound to the mutation site; and (d) comparing the chromatogram of step (c) to the chromatogram of step (a), wherein a change in the retention time or the number of peaks or other shapes in the chromatogram of step (c) indicates the presence of a mutation in said sample. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
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Specification