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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. doi:10.1126/science.2814502.

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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
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URLURL TypeDescription 10.1126/science.2814502 DOIArticle JSTORArticle
Wold, Barbara0000-0003-3235-8130
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.).
Funding AgencyGrant Number
National Institute of AgingAG08196
Deutsche Forschungsgemeinschaft (DFG)Pf212/1-1
Issue or Number:4931
Record Number:CaltechAUTHORS:20150203-134224388
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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
Deposited On:04 Feb 2015 00:07
Last Modified:10 Nov 2021 20:32

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