LIGATION-MEDIATED PCR FOR CHROMATIN-STRUCTURE ANALYSIS OF INTERPHASE AND METAPHASE CHROMATIN

Chromatin structure is becoming increasingly recognized as important f or a full understanding of gene function and cell memory. With regard to cell memory, which involves the transfer of chromatin-encoded epige netic information from one cell generation to another, the detailed st ructure of metaphase chromatin is of crucial importance. In this paper we describe methods for the use of dimethyl sulfate, DNase I, and pot assium permanganate for in vivo footprinting and chromatin analysis, w ith 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 studie s of a matrix attachment region near the human beta-interferon gene. ( C) 1997 Academic Press.