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Genomic Sequencing and Methylation Analysis by Ligation Mediated PCR

Pfeiffer, Gerd P. and Steigerwald, Sabine D. and Mueller, Paul R. and Wold, Barbara and Riggs, Arthur D. (1989) Genomic Sequencing and Methylation Analysis by Ligation Mediated PCR. Science, 246 (4931). pp. 810-813. ISSN 0036-8075. http://resolver.caltech.edu/CaltechAUTHORS:20150203-134224388

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Abstract

Genomic sequencing permits studies of in vivo DNA methylation and protein-DNA interactions, but its use has been limited because of the complexity of the mammalian genome. A newly developed genomic sequencing procedure in which a ligation mediated polymerase chain reaction (PCR) is used generates high quality, reproducible sequence ladders starting with only 1 microgram of uncloned mammalian DNA per reaction. Different sequence ladders can be created simultaneously by inclusion of multiple primers and visualized separately by rehybridization. Relatively little radioactivity is needed for hybridization and exposure times are short. Methylation patterns in genomic DNA are readily detectable; for example, 17 CpG dinucleotides in the 5' region of human X-linked PGK-1 (phosphoglycerate kinase 1) were found to be methylated on an inactive human X chromosome, but unmethylated on an active X chromosome.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/ 10.1126/science.2814502 DOIArticle
http://www.sciencemag.org/content/246/4931/810PublisherArticle
http://www.jstor.org/stable/1704730 JSTORArticle
Additional Information:© 1989 American Association for the Advancement of Science. 11 July 1989; accepted2 6 September 1989. Supported by National Institute of Aging grant AG08196 (A.D.R.), NIH grants RR07003 and GM355262BW (B.W.), and a fellowship from the Deutsche Forschungsgemeinschaft (Pf212/1-1) (G-P.P.).
Funders:
Funding AgencyGrant Number
National Institute of AgingAG08196
NIHRR07003
NIHGM355262BW
Deutsche Forschungsgemeinschaft (DFG)Pf212/1-1
Record Number:CaltechAUTHORS:20150203-134224388
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150203-134224388
Official Citation:Pfeifer, G., Steigerwald, S., Mueller, P., Wold, B., & Riggs, A. (1989). Genomic sequencing and methylation analysis by ligation mediated PCR. Science, 246(4931), 810-813. doi: 10.1126/science.2814502
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:54330
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:04 Feb 2015 00:07
Last Modified:04 Feb 2015 00:07

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