Method for detecting cytosine methylations

Details Method for detecting cytosine methylations Method for detecting cytosine methylations

2006-10-10

2006-10-10

Martinell, James (Department: 1634)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

C435S091100, C435S091200

Reexamination Certificate

active

07118868

ABSTRACT:
A method is described for the detection of 5-methylcytosine in genomic DNA samples. First, a genomic DNA from a DNA sample is chemically converted with a reagent, whereby 5-methylcytosine and cytosine react differently. Then the pretreated DNA is amplified with the use of a polymerase with primers of different sequence. In the next step, the amplified genomic DNA is hybridized to an oligonucleotide array and PCR products are obtained, which must be provided with a label. Alternatively, the PCR products can be extended in a primer extension reaction, wherein the extension products are also provided with a label.In the last step, the extended oligonucleotides are investigated for the presence of the label.

REFERENCES:
patent: WO 00/31294 (2000-06-01), None
Oxis Research (http://www.oxisresearch.com/products/chemicals/peroxynitrite/33113/33113.shtml (Jul. 19, 2005).
Herman et al, Proc. Natl. Acad. Sci. USA 93: 9821 (1996).
USB Molecular Biology Reagents/Protocols 1992, United States Biochemical Corporation, 1992, Cleveland, OH, pp. 218-219.
Xiong et al., ‘COBRA: a sensitive and quantitative DNA methylation assay.’ Nucleic Acids Research, 25(12):2532-4 (1997).

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