Skip Navigation

Nucleic Acids Research 2004 32(21):e167; doi:10.1093/nar/gnh165
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (694K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Schatz, P.
Right arrow Articles by Schuster, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schatz, P.
Right arrow Articles by Schuster, M.
Related Collections
Right arrow Nucleic acid modification
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 2 December 2004

Nucleic Acids Research, Vol. 32 No. 21 © Oxford University Press 2004; all rights reserved

Rapid analysis of CpG methylation patterns using RNase T1 cleavage and MALDI-TOF

Philipp Schatz, Dimo Dietrich and Matthias Schuster*

Epigenomics AG, Science Department, Kleine Präsidentenstraße 1, D-10178 Berlin, Germany

* To whom correspondence should be addressed. Tel: +49 30 24345100; Fax: +49 30 24345299; Email: schuster{at}epigenomics.com
The authors wish to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Received July 2, 2004; Revised October 11, 2004; Accepted November 4, 2004

Here, we introduce a method for the fast and accurate analysis of DNA methylation based on bisulfite-treated DNA. The target region is PCR amplified using a T7 RNA polymerase promoter-tagged primer. A subsequent in vitro transcription leads to a transcript which contains guanosine residues only at sites that contained methylated cytosines before bisulfite treatment. In a single tube reaction using guanosine-specific cleavage by RNase T1, a specific pattern of RNA fragments is formed. This pattern directly represents the methylation state of the sample DNA and is analyzed using matrix-assisted laser desorption ionization time-of-flight technology. This method was successfully applied to the analysis of artificially methylated and unmethylated DNA, mixtures thereof and colon DNA samples. The applicability for the analysis of both PCR products and cloned PCR products is demonstrated. The observed methylation patterns were confirmed by bisulfite sequencing.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 DevelopmentHome page
D. Zilberman and S. Henikoff
Genome-wide analysis of DNA methylation patterns
Development, November 15, 2007; 134(22): 3959 - 3965.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. W. Coolen, A. L. Statham, M. Gardiner-Garden, and S. J. Clark
Genomic profiling of CpG methylation and allelic specificity using quantitative high-throughput mass spectrometry: critical evaluation and improvements
Nucleic Acids Res., September 25, 2007; 35(18): e119 - e119.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Pfister, C. Schlaeger, F. Mendrzyk, A. Wittmann, A. Benner, A. Kulozik, W. Scheurlen, B. Radlwimmer, and P. Lichter
Array-based profiling of reference-independent methylation status (aPRIMES) identifies frequent promoter methylation and consecutive downregulation of ZIC2 in pediatric medulloblastoma
Nucleic Acids Res., April 1, 2007; 35(7): e51 - e51.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
K. Brakensiek, L. U. Wingen, F. Langer, H. Kreipe, and U. Lehmann
Quantitative High-Resolution CpG Island Mapping with PyrosequencingTM Reveals Disease-Specific Methylation Patterns of the CDKN2B Gene in Myelodysplastic Syndrome and Myeloid Leukemia
Clin. Chem., January 1, 2007; 53(1): 17 - 23.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Fu, S. Xu, C. Pan, M. Ye, H. Zou, and B. Guo
A matrix of 3,4-diaminobenzophenone for the analysis of oligonucleotides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Nucleic Acids Res., August 2, 2006; 34(13): e94 - e94.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
M. Procter, L.-S. Chou, W. Tang, M. Jama, and R. Mao
Molecular Diagnosis of Prader-Willi and Angelman Syndromes by Methylation-Specific Melting Analysis and Methylation-Specific Multiplex Ligation-Dependent Probe Amplification
Clin. Chem., July 1, 2006; 52(7): 1276 - 1283.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
S. Ogino, T. Kawasaki, M. Brahmandam, M. Cantor, G. J. Kirkner, D. Spiegelman, G. M. Makrigiorgos, D. J. Weisenberger, P. W. Laird, M. Loda, et al.
Precision and Performance Characteristics of Bisulfite Conversion and Real-Time PCR (MethyLight) for Quantitative DNA Methylation Analysis
J. Mol. Diagn., May 1, 2006; 8(2): 209 - 217.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
P. Schatz, J. Distler, K. Berlin, and M. Schuster
Novel method for high throughput DNA methylation marker evaluation using PNA-probe library hybridization and MALDI-TOF detection.
Nucleic Acids Res., January 1, 2006; 34(8): e59 - e59.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
I. Yang, I. Y. Park, S.-M. Jang, L. H. Shi, H.-K. Ku, and S.-R. Park
Rapid quantification of DNA methylation through dNMP analysis following bisulfite-PCR.
Nucleic Acids Res., January 1, 2006; 34(8): e61 - e61.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.