Methods for detection of methylated DNA
First Claim
1. A method for preparing DNA template for DNA methylation analysis, the method comprising incubating DNA template at a high temperature in the presence of a bisulfite reagent to produce the combined denaturation and bisulfite conversion of the DNA template.
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Abstract
The present invention provides methods for improving the efficiency of methylation detection. The present invention provides improvements from current technologies via increased efficiency of template denaturation and bisulfite conversion reaction, also significant time savings in sample preparation, recovery, as well as increased efficiency of desulfonation. These methods facilitate rapid analysis of research and clinical samples and enhance the ability to process high-through put sample preparations. The methods are applicable to essentially all methylation detection procedures and also to the analysis of methylation patterns from various species.
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Citations
32 Claims
- 1. A method for preparing DNA template for DNA methylation analysis, the method comprising incubating DNA template at a high temperature in the presence of a bisulfite reagent to produce the combined denaturation and bisulfite conversion of the DNA template.
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16. A method for preparing denatured, bisulfite converted, and desulfonated DNA template in a coupled process, the method comprising:
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a) denaturating and bisulfite converting DNA at a temperature from about 90°
C. to about 100°
C.;
b) adsorbing the DNA to a solid matrix;
c) desulfonating the adsorbed DNA; and
d) eluting the desulfonated DNA from the solid matrix. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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30. A column for adsorbing DNA for combined denaturation and bisulfite conversion at a high temperature coupled to desulfonation, the column having a cylindrical bed comprising a silica resin, providing for elution of adsorbed DNA in an aqueous solution.
- 31. A kit of parts for preparing DNA template for DNA methylation analysis, the kit comprising a dry bisulfite reagent and solutions for combined denaturation and bisulfite conversion at a high temperature and a solid matrix for binding DNA during desulfonation and for eluting the template DNA produced.
Specification