An Improved Restriction Enzyme Accessibility Assay for Analyzing Changes in Chromatin Structure in Samples of Limited Cell Number
UMass Chan Affiliations
Department of Cell BiologyDocument Type
Book ChapterPublication Date
2012-01-01Keywords
DNA Restriction EnzymesChromatin Assembly and Disassembly
Muscle, Skeletal
Enzyme Assays
Cell Biology
Metadata
Show full item recordAbstract
Studies investigating mechanisms that control gene regulation frequently examine the accessibility of specific DNA sequences to nuclease cleavage. In general, sequences that are sensitive to nuclease cleavage are considered to be in an "open" chromatin conformation that is associated with regulatory factor binding, while sequences resistant to nuclease cleavage are considered to be in a "closed" conformation commonly associated with chromatin that is neither poised for transcription nor being actively transcribed. Changes in nuclease accessibility at specific genomic sequences reflect changes in the local chromatin structure that can occur as a result of signaling cues in the extracellular environment. These changes in chromatin structure usually precede or are coincident with changes in gene expression patterns and are therefore a useful marker of regulatory events controlling transcription. We describe a method to perform restriction enzyme accessibility assays (REAA) that utilizes ligation-mediated polymerase chain reaction (LM-PCR) technology and that permits assessment of samples from any source containing as few as 1,000 cells. Use of this modified REAA protocol will enhance analysis of chromatin structural changes at specific DNA sequences of interest by making it possible to analyze samples where unrestricted amounts of sample are not readily available.Source
Methods Mol Biol. 2012;798:531-42. Link to article on publisher's siteDOI
10.1007/978-1-61779-343-1_32Permanent Link to this Item
http://hdl.handle.net/20.500.14038/34852PubMed ID
22130859Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1007/978-1-61779-343-1_32