Ligation-mediated PCR for chromatin-structure analysis of interphase and metaphase chromatin.
Chromatin structure is becoming increasingly recognized as important for a full understanding of gene function and cell memory. With regard to cell memory, which involves the transfer of chromatin-encoded epigenetic information from one cell generation to another, the detailed structure of metaphase chromatin is of crucial importance. In this paper we describe methods for the use of dimethyl sulfate, DNase I, and potassium permanganate for in vivo footprinting and chromatin analysis, with special emphasis on studies of metaphase cells. We review the use of ligation-mediated PCR for the analysis of chromatin, including the human phosphoglycerate kinase promoter, and also report initial studies of a matrix attachment region near the human beta-interferon gene.
Hershkovitz
M
M
Biology Department, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
Riggs
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AD
D002460:Cell Line; D002843:Chromatin / Q000502:physiology / Q000648:ultrastructure*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D018983:DNA Footprinting / Q000379:methods*; D019175:DNA Methylation; D017931:DNA Primers; D015342:DNA Probes; D003850:Deoxyribonuclease I; D005347:Fibroblasts; D006801:Humans; D007202:Indicators and Reagents; D007231:Infant, Newborn; D016899:Interferon-beta / Q000096:biosynthesis / Q000235:genetics; D007399:Interphase; D008297:Male; D008677:Metaphase; D016133:Polymerase Chain Reaction / Q000379:methods*; D012045:Regulatory Sequences, Nucleic Acid; D013463:Sulfuric Acid Esters; D001749:Urinary Bladder Neoplasms
D002843:Chromatin; D017931:DNA Primers; D015342:DNA Probes; D007202:Indicators and Reagents; D013463:Sulfuric Acid Esters; D016899:Interferon-beta; D004247:DNA; D003850:Deoxyribonuclease I; C007482:dimethyl sulfate