Abstract
In mammals, methylation of cytosine C-5 position is a major heritable epigenetic mark on the DNA molecule. Maintenance of proper DNA methylation patterns is a key process during embryo development and in the maintenance of adult tissue homeostasis. The use of experimental procedures based on the chemical modification of cytosine by sodium bisulfite and the development of antibodies recognizing 5mC have essentially contributed to our knowledge on DNA methylation dynamics in normal and disease states. Here we describe standard procedures for bisulfite sequencing, methylation-specific PCR, and 5mC immunodetection using mouse skin and the hair follicle stem cell niche as model tissues.
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Acknowledgements
This work was supported by grants of the Spanish Ministerio de Economía y Competitividad (SAF 11-23493) and the Comunidad Autónoma de Madrid (SkinModel, CAM S10/BMD-2359) to J.E.; E.C. and M.I.C. are supported by PhD fellowship grants of the Spanish Ministerio de Educación and Universidad Autónoma de Madrid, respectively.
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Espada, J., Carrasco, E., Calvo, M.I. (2014). Standard DNA Methylation Analysis in Mouse Epidermis: Bisulfite Sequencing, Methylation-Specific PCR, and 5-Methyl-Cytosine (5mC) Immunological Detection. In: Stockert, J., Espada, J., Blázquez-Castro, A. (eds) Functional Analysis of DNA and Chromatin. Methods in Molecular Biology, vol 1094. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-706-8_17
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DOI: https://doi.org/10.1007/978-1-62703-706-8_17
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Publisher Name: Humana Press, Totowa, NJ
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