Direct Sensing of 5-Methylcytosine by Polymerase Chain Reaction
Direct Sensing of 5-Methylcytosine by Polymerase Chain Reaction
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2014
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339834
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EvoEPIGEN - Evolved Replication Systems of Epigenetics
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Angewandte Chemie International Edition ; 53 (2014), 31. - pp. 8154-8158. - ISSN 1433-7851. - eISSN 1521-3773
Abstract
The epigenetic control of genes by the methylation of cytosine resulting in 5-methylcytosine (5mC) has fundamental implications for human development and disease. Analysis of alterations in DNA methylation patterns is an emerging tool for cancer diagnostics and prognostics. Here we report that two thermostable DNA polymerases, namely the DNA polymerase KlenTaq derived from Thermus aquaticus and the KOD DNA polymerase from Thermococcus kodakaraensis, are able to extend 3′-mismatched primer strands more efficiently from 5 mC than from unmethylated C. This feature was advanced by generating a DNA polymerase mutant with further improved 5mC/C discrimination properties and its successful application in a novel methylation-specific PCR approach directly from untreated human genomic DNA.
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540 Chemistry
Keywords
DNA methylation,DNA polymerases,enzyme engineering,polymerase chain reaction
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ASCHENBRENNER, Joos, Matthias DRUM, Hüsnü TOPAL, Markus WIELAND, Andreas MARX, 2014. Direct Sensing of 5-Methylcytosine by Polymerase Chain Reaction. In: Angewandte Chemie International Edition. 53(31), pp. 8154-8158. ISSN 1433-7851. eISSN 1521-3773. Available under: doi: 10.1002/anie.201403745BibTex
@article{Aschenbrenner2014-07-28Direc-28340, year={2014}, doi={10.1002/anie.201403745}, title={Direct Sensing of 5-Methylcytosine by Polymerase Chain Reaction}, number={31}, volume={53}, issn={1433-7851}, journal={Angewandte Chemie International Edition}, pages={8154--8158}, author={Aschenbrenner, Joos and Drum, Matthias and Topal, Hüsnü and Wieland, Markus and Marx, Andreas} }
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