Simple screening method for differentially methylated regions of the genome using a small number of cells.
Genomic DNA methylation is a major epigenetic mechanism controlling the expression of genetic information. Therefore, identifying regions of the genome that are differentially methylated in different cells is a useful strategy for the study of biological phenomena. To date, several useful screening methods have been established for identifying differentially methylated genomic regions. However, it is impossible to use these methods in fields of study in which it is difficult to obtain a large number of uniform cells, because considerable amounts of genomic DNA are required. Given this situation, we developed a method for preparing large genomic DNA from a small number of cells, and a simple and highly sensitive method for screening for differentially methylated sites. Combined, these two methods comprise a simple screening method, which we named "Differential Methylation Site Scanning" (DMSS), for identifying differentially methylated regions of the genome from a small number of cells. Just 10 cells are sufficient for the method described here.
Hamada
Tsuyoshi
T
Department of Anatomy and Neurobiology, National Defense Medical Collage, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan. thamada@ndmc.ac.jp
Murasawa
Satoshi
S
Asahara
Takayuki
T
D002460:Cell Line; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D019175:DNA Methylation; D006367:HeLa Cells; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004247:DNA