Abstract
A novel procedure for DNA methylation analysis is described that characterizes the extent of DNA methylation in CpG islands. The basic concept relies on direct immunodetection of 5′ methylcytosines (5′ mCs) without the need for bisulfite treatment utilizing a microarray format. This system is designed for the application of immunofluorescence using a monoclonal antibody that specifically recognizes 5′ mC in single-stranded DNA hybridized to oligonucleotide microarrays. An ultrasensitive fluorescence scanner and 170−μm thin aldehyde-functionalized glass slides are used to optimize the signal-to-noise ratio and to minimize autofluorescence. These methodological improvements allow for the direct detection of 5′ mC in genomic DNA hybridized to microarrays without prior PCR amplification with high analytical sensitivity.
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References
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Acknowledgements
This study was supported by the federal government of Upper Austria and by the Austrian Federal Ministry for Education, Science and Culture in the context of the Gen-AU project “Ultra-sensitive Proteomics and Genomics”.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Pröll, J., Wechselberger, C., Födermayr, M., Zach, O., Lutz, D. (2009). Immunodetection Array. In: Tost, J. (eds) DNA Methylation. Methods in Molecular Biology, vol 507. Humana Press. https://doi.org/10.1007/978-1-59745-522-0_4
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DOI: https://doi.org/10.1007/978-1-59745-522-0_4
Publisher Name: Humana Press
Print ISBN: 978-1-934115-61-9
Online ISBN: 978-1-59745-522-0
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