Nucleic acid detection assay
First Claim
1. A method for determining the methylation status of a potential methylation site in genomic nucleic acid comprising:
- treating genomic nucleic acid with an agent which modifies cytosine bases but does not modify 5-methyl-cytosine bases under conditions to form a modified nucleic acid template from two complementary strands of genomic nucleic acid containing a potential methylation site;
providing a first clamp containing a first capture sequence and a second capture sequence, first capture sequence being complementary to a region flanking one side of the potential methylation site in the first strand of the modified nucleic acid template, and the second capture sequence being complementary to a region flanking one side of the potential methylation site in the second strand of the modified nucleic acid template;
providing a second clamp containing a third capture sequence complementary to a region flanking the other side of the potential methylation site in the modified nucleic acid template;
allowing the first clamp and the second clamp to hybridise to the modified nucleic acid template;
ligating the hybridised first and second clamps to form a probe spanning the potential methylation site in the modified nucleic acid template;
digesting the modified nucleic acid template to obtain the probe; and
detecting the probe and determining the methylation status of the potential methylation site in the modified genomic nucleic acid.
0 Assignments
0 Petitions
Accused Products
Abstract
A method for determining the methylation status of a potential methylation site in genomic nucleic acid comprising treating genomic nucleic acid with an agent which modifies cytosine bases but does not modify 5methyl-cytosine bases under conditions to form a. modified nucleic acid template containing a potential methylation site; providing a first clamp containing a first capture sequence complementary to a region flanking one side of the potential methylation site in the modified nucleic acid template, providing a second clamp containing a second capture sequence complementary to a region flanking the other side of the potential methylation site in the modified nucleic acid template; allowing the first clamp and the second clamp to hybridise to the modified nucleic acid template; ligating the hybridised first and second clamps to form a probe spanning the potential methylation site in the modified nucleic acid template; digesting the modified nucleic acid template to obtain the probe; and detecting the probe and determining the methylation status of the potential methylation site in the modified genomic nucleic acid.
-
Citations
41 Claims
-
1. A method for determining the methylation status of a potential methylation site in genomic nucleic acid comprising:
-
treating genomic nucleic acid with an agent which modifies cytosine bases but does not modify 5-methyl-cytosine bases under conditions to form a modified nucleic acid template from two complementary strands of genomic nucleic acid containing a potential methylation site;
providing a first clamp containing a first capture sequence and a second capture sequence, first capture sequence being complementary to a region flanking one side of the potential methylation site in the first strand of the modified nucleic acid template, and the second capture sequence being complementary to a region flanking one side of the potential methylation site in the second strand of the modified nucleic acid template;
providing a second clamp containing a third capture sequence complementary to a region flanking the other side of the potential methylation site in the modified nucleic acid template;
allowing the first clamp and the second clamp to hybridise to the modified nucleic acid template;
ligating the hybridised first and second clamps to form a probe spanning the potential methylation site in the modified nucleic acid template;
digesting the modified nucleic acid template to obtain the probe; and
detecting the probe and determining the methylation status of the potential methylation site in the modified genomic nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 41)
-
-
21. A method for determining the methylation status of a potential methylation site on genomic nucleic acid comprising:
-
treating genomic nucleic acid with an agent which modifies cytosine bases but does not modify 5-methyl-cytosine bases under conditions to form a modified nucleic acid template from two complementary strands of genomic nucleic acid containing a potential methylation site;
providing a first clamp containing a first capture sequence and a second capture sequence, first capture sequence being complementary to a region flanking one side of the potential methylation site in the first strand of the modified nucleic acid template, and the second capture sequence being complementary to a region flanking one side of the potential methylation site in the second strand of the modified nucleic acid template;
providing a second clamp containing a third capture sequence and a fourth capture sequence, the third capture sequence being complementary to a region flanking the other side the potential methylation site in the first strand of the modified nucleic acid template, and the fourth capture sequence being complementary to a region flanking the other side of the potential methylation site in the second strand of the modified nucleic acid template;
allowing the first clamp and the second clamp to hybridise to the two complementary strands of the modified nucleic acid template;
ligating the hybridised clamps to form a circular probe spanning the potential methylation site in the complementary strands of the modified nucleic acid template;
digesting the modified nucleic acid template to obtain the circular probe; and
detecting the circular probe and determining the methylation status of the potential methylation site in the modified genomic nucleic acid. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
-
Specification