Standard DNA methylation analysis in mouse epidermis: Bisulfite sequencing, methylation-specific PCR, and 5-methyl-cytosine (5mC) immunological detection

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 che...

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Detalles Bibliográficos
Autores: Espada, Jesús, Carrasco, Elisa, Calvo, María I.
Tipo de recurso: otro
Estado:Versión enviada para evaluación y publicación
Fecha de publicación:2014
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/110701
Acceso en línea:http://hdl.handle.net/10261/110701
Access Level:acceso abierto
Palabra clave:DNA methylation
5-Methyl-cytosine (5mC9)
Mouse epidermis
Hair follicle
Bisulfite sequencing
Immunological detection
Descripción
Sumario: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.