Restriction endonuclease accessibility as a determinant of altered chromatin structure.
Active genes in eukaryotic genomes are typically found in open, nuclease-sensitive regions of chromatin. This chapter presents an overview of the techniques used to assay restriction endonuclease cleavage of chromosomal DNA as an approach to assess general accessibility at a genomic region of interest. We describe protocols to (1) prepare nuclei templates, (2) treat chromosomal DNA with a restriction enzyme(s), and (3) visualize and quantify chromosomal cleavage(s), with an emphasis on ligation-mediated (LM) PCR techniques.
Department of Research Technologies, Pfizer Global Research and Development, Fresnes Cedex, France.
William M
WM
Hempel
Pierre
P
Ferrier
D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry; D002469:Cell Separation / Q000379:methods; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping / Q000379:methods*
D002843:Chromatin; D004247:DNA; D004262:DNA Restriction Enzymes