Genomic sequencing by ligation-mediated PCR |
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Authors: | Gerd P. Pfeifer Arthur D. Riggs |
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Affiliation: | (1) Department of Biology, Beckman Research Institute of the City of Hope, 91010 Duarte, CA |
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Abstract: | Genomic sequencing permits studies of in vivo DNA methylation and protein-DNA interactions, but its use has been limited due to the complexity of the mammalian genome. Ligation-mediated PCR (LMPCR) is a sensitive genomic sequencing procedure that generates high quality, reproducible sequence ladders starting with only 1 μg of uncloned mammalian DNA per reaction. This genomic sequencing procedure can be adapted for various methylation, in vivo footprinting and DNA adduct mapping procedures. We provide a detailed protocol for genomic sequencing by LMPCR and discuss the principles and applications of the method. |
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Keywords: | Chromatin structure gene regulation polymerase chain reaction DNA methylation in vivo footprinting mutagenesis |
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