Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2005-12-20
2005-12-20
Fredman, Jeffrey (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091100, C435S091200, C536S024300, C536S023100
Reexamination Certificate
active
06977146
ABSTRACT:
The invention concerns a method for the identification of cytosine methylation patterns in genomic DNA samples, whereina) a genomic DNA sample is chemically treated such that cytosine and 5-methylcytosine react differently and a different base pairing behavior of the two products results in the duplex;b) parts of the thus-treated DNA sample are enzymatically amplified;c) the amplified parts of the thus-treated DNA sample bind to a surface;d) a set of probes of different nucleobase sequences, each of which contains the dinucleotide sequence 5′-CpG-3′ at least once, is hybridized to the immobilized DNA sample;e) the non-hybridized probes are separated;f) the hybridized probes are analyzed in a mass spectrometer, wherein the position of the probes on the sample holder permits a classification of the hybridizing DNA sample;g) Assignment of the peak pattern obtained from the mass spectra to the methylation pattern and comparison of the new data with a database.
REFERENCES:
patent: 5691141 (1997-11-01), Koster
patent: 5786146 (1998-07-01), Herman et al.
patent: 6265171 (2001-07-01), Herman et al.
Katouzian-Safadi et al., “Identification of the DNA-interacting sites of proteins: Micro sequencing of the peptides cross-linked to 5-bromouracil substituted DNA”, Biochimie, (1994), vol. 76 (2), pp. 129-132.
Herman et al., “Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands,” Proc. Natl. Acad. Sci. USA, 93:9821-6 (Sep. 1996).
Frommer et al., “A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands,” Proc. Natl. Acad. Sci. USA, 89:1827-31.
Epigenomics AG
Fredman Jeffrey
Kriegsman & Kriegsman
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