System and method for temperature gradient capillary electrophoresis
First Claim
1. A system for determining the presence of a mutation in at least one sample polynucleotide, comprising:
- a processor configured to at least;
receive (1) at least a first set of sample electrophoresis data indicative of a temperature gradient electrophoresis (TGE) separation of at least one sample polynucleotide, and (2) at least a first set of reference electrophoresis data indicative of a TGE separation of at least one reference polynucleotide; and
process at least a subset of the first set of sample electrophoresis data and at least a subset of the first set of reference electrophoresis data to prepare a set of result data corresponding to the first set of sample electrophoresis data, wherein the set of result data includes data indicative of whether a sample polynucleotide of the first set of sample electrophoresis data includes a mutation.
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Abstract
The present invention relates to a method for determining the presence of a mutation in a first sample comprising first polynucleotides. The reference sample comprises reference polynucleotides. The first sample and a reference sample are subjected to electrophoresis in the presence of at least one intercalating dye. During electrophoresis the temperature of the first sample and the reference sample is changed by an amount sufficient to change an electrophoretic mobility of at least one of the first or reference polynucleotides. Fluorescence intensity data are obtained. The fluorescence intensity data are indicative of the presence of the first and reference polynucleotides. The data are processed to determine the presence of mutation in the first polynucleotides.
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Citations
31 Claims
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1. A system for determining the presence of a mutation in at least one sample polynucleotide, comprising:
a processor configured to at least;
receive (1) at least a first set of sample electrophoresis data indicative of a temperature gradient electrophoresis (TGE) separation of at least one sample polynucleotide, and (2) at least a first set of reference electrophoresis data indicative of a TGE separation of at least one reference polynucleotide; and
process at least a subset of the first set of sample electrophoresis data and at least a subset of the first set of reference electrophoresis data to prepare a set of result data corresponding to the first set of sample electrophoresis data, wherein the set of result data includes data indicative of whether a sample polynucleotide of the first set of sample electrophoresis data includes a mutation. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A computer-readable medium comprising executable software code, the code for determining the presence of a mutation in at least one sample polynucleotide, comprising:
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code to receive (1) at least a first set of sample electrophoresis data indicative of a temperature gradient electrophoresis (TGE) separation of at least one sample polynucleotide, and (2) at least a first set of reference electrophoresis data indicative of a TGE separation of at least one reference polynucleotide; and
code to process at least a subset of the first set of sample electrophoresis data and at least a subset of the first set of reference electrophoresis data to prepare a set of result data corresponding to the first set of sample electrophoresis data, wherein the set of result data includes data indicative of whether a sample polynucleotide of the first set of sample electrophoresis data includes a mutation. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. A method for interacting with a computer to determine the presence of mutation in at least one sample polynucleotide, comprising the steps of:
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executing an application which includes one or more windows having one or more controls;
manipulating at least one of said controls to define members of a group of sample electrophoresis data;
manipulating at least one of said controls for defining a set of reference electrophoresis data to be used for processing members of the group;
manipulating at least one of said controls to select a first subset of each member of the group and a first subset of the reference electrophoresis data; and
manipulating at least one of said controls to execute code configured to process the first subsets of the members of the group and the first subset of the set of reference electrophoresis data to prepare result data corresponding to each member of the group, wherein the result data corresponding to each member of the group includes data indicative of whether a first sample polynucleotide of the corresponding sample electrophoresis data includes a mutation.
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26. The method of claim 25, further comprising the steps of:
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manipulating at least one of said controls to select a second subset of each member of the group and a second subset of the reference electrophoresis data; and
manipulating at least one of said controls to execute code configured to process the second subsets of the members of the group and the second subset of the set of reference electrophoresis data to prepare result data corresponding to each member of the group, wherein the result data corresponding to each member of the group includes data indicative of whether a second sample polynucleotide of the corresponding sample electrophoresis data includes a mutation. - View Dependent Claims (27, 28, 29, 30)
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26-1. A method of processing electrophoresis data obtained by temperature gradient electrophoresis (TGE) separation to provide data indicative of the presence of a single polynucleotide polymorphism (SNP) or a mutation in a sample compound of a biological sample, comprising:
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providing reference electrophoresis data comprising a plurality of reference data points (dri, mi), the reference data points defining at least one reference peak and having been obtained by TGE of at least one reference compound, and where, for the ith reference data point, dri is a detection value indicative of a detector signal and mi is a migration coordinate;
providing sample electrophoresis data comprising a plurality of sample data points (dsj, mj), the sample data points defining at least one sample peak and having been obtained by TGE of at least one sample compound, and where, for the jth sample data point, dsj is a detection value indicative of a detector signal and mj is a migration coordinate;
normalizing the reference and sample electrophoresis data based on at least one detection value of the reference electrophoresis data and at least one detection value of the sample electrophoresis data;
determining a plurality of detection value differences Δ
between the reference and sample electrophoresis data, where the kth detection value difference Δ
k is indicative of a difference between (1) the detection value dri of the reference electrophoresis data point having the migration coordinate mi and (2) the detection value dsj of the sample electrophoresis data point having the migration coordinate mj; and
wherein the plurality of detection value differences Δ
are indicative of the presence of an SNP or mutation in the sample compound of the biological sample.
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31. A method of processing electrophoresis data obtained by temperature gradient electrophoresis (TGE) separation to provide data indicative of the presence of a single polynucleotide polymorphism (SNP) or a mutation in a sample compound of a biological sample, comprising:
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providing reference electrophoresis data comprising a plurality of reference data points (dri, mi), the reference data points defining at least one reference peak and having been obtained by TGE of at least one reference compound, and where, for the ith reference data point, dsi is a detection value indicative of a detector signal and mi is a migration coordinate;
providing sample electrophoresis data comprising a plurality of sample data points (sj, mj), the sample data points defining at least one sample peak and having been obtained by TGE of at least one sample compound, and where, for the jth sample data point, sj is a detection value indicative of a detector signal and mj is a migration coordinate;
normalizing the reference and sample electrophoresis data based on at least one detection value of the reference electrophoresis data and at least one detection value of the sample electrophoresis data; and
determining a covariance between the reference and sample electrophoresis data, wherein the covariance is indicative of the presence of an SNP or mutation in the sample compound of the biological sample.
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Specification