title: DNA methylation profiling of hematopoietic stem cells.
abstract: DNA methylation is a key epigenetic mark that is essential for properly functioning hematopoietic stem cells. Determining where functionally relevant DNA methylation marks exist in the genome is crucial to understanding the role that methylation plays in hematopoiesis. This chapter describes a method to profile DNA methylation by selectively enriching methylated DNA sequences that are bound in vitro by methyl-binding domain (MBD) proteins. The MBD-pulldown approach selects for DNA sequences that have the potential to be "read" by the endogenous machinery involved in epigenetic regulation. Furthermore, this approach is feasible with very small quantities of DNA, and is compatible with the use of any downstream high-throughput sequencing approach. This technique offers a reliable, simple, and powerful tool for exploration of the role of DNA methylation in hematopoietic stem cells.
authors: Division of Human Genetics, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, College of Medicine, University of Cincinnati, 3333 Burnet Avenue, Cincinnati, OH, 45229, USA, amber.begtrup@cchmc.org.
authors: Amber Hogart
authors: AH
authors: Begtrup
mesh_terms: D000818:Animals; D002469:Cell Separation; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism; D023281:Genomics; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D011786:Quality Control
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA