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Nucleic acid detection assay

  • US 20070178459A1
  • Filed: 09/03/2004
  • Published: 08/02/2007
  • Est. Priority Date: 09/04/2003
  • Status: Active Grant
First Claim
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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.

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