Title:
DNA中のシトシンのメチル化法および検出法
Document Type and Number:
Japanese Patent JP4743787
Kind Code:
B2
Abstract:
Detecting cytosine methylation in DNA (I) by reacting (I) with an enzyme or chemical so that 5-methylcytosine (5MC) remains unchanged, while non-methylated cytosine (C) is converted to uracil or some other base having base-pairing properties different from those of C, is new. Detecting cytosine methylation in DNA (I) comprises: (a) reacting (I) with an enzyme or chemical so that 5-methylcytosine (5MC) remains unchanged, while non-methylated cytosine (C) is converted to uracil or some other base having base-pairing properties different from those of C; (b) amplifying the pretreated (I) using a polymerase and at least one primer that is linked at its 5'-end to a probe (scorpion primer, SP); (c) separating the primer extension product (PEP) from the matrix strand; (d) intracellular hybridization of the probe to PEP, where hybridization is dependent on the methylation status of the DNA; and (e) detecting whether or not hybridization of the probe has occurred. An independent claim is also included for a kit comprising SP, polymerase and reagents for performing a PCR.
Inventors:
Tezner, Laimo
Distler, Jurgen
Distler, Jurgen
Application Number:
JP2006523519A
Publication Date:
August 10, 2011
Filing Date:
August 13, 2004
Export Citation:
Assignee:
Epigenomics Age
International Classes:
C12Q1/68; C12N15/09
Foreign References:
| US20030087240 | ||||
| US6365729 | ||||
| US6326145 |
Other References:
Nucleic acids research. 2000, Vol.28, No.19, p.3752-3761
Nucleic acids research. 2001, Vol.29, No.20, e96(p.1-9)
Proceedings of the National Academy of Sciences of the United States of America. 1996, Vol.93, No.18, p.9821-9826
Nucleic acids research. 2001, Vol.29, No.20, e96(p.1-9)
Proceedings of the National Academy of Sciences of the United States of America. 1996, Vol.93, No.18, p.9821-9826
Attorney, Agent or Firm:
Eiichiro Shimazaki
Kanji Arai
Naotaka Kono
Kanji Arai
Naotaka Kono