title: Standard DNA methylation analysis in mouse epidermis: bisulfite sequencing, methylation-specific PCR, and 5-methyl-cytosine (5mC) immunological detection.
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.
authors.affiliation: Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Espada
authors.forename: Elisa
authors.initials: E
authors.lastname: Carrasco
authors.forename: María I
authors.initials: MI
authors.lastname: Calvo
mesh_terms: D044503:5-Methylcytosine / Q000276:immunology*; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification; D019175:DNA Methylation; D004817:Epidermis / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000737:chemistry*; D013623:Tail
chemical_list: D013447:Sulfites; D044503:5-Methylcytosine; D004247:DNA; C042345:hydrogen sulfite