Abstract
Publisher Summary This chapter focuses on the reconstitution of chromatin from purified components. Several useful methods have resulted from attempts to reconstitute purified histories and DNA into chromatin in vitro. Alternatively, core histories can be added very slowly to DNA at physiological salt concentrations to form nucleosomes. Both of these methods have been devised to circumvent the aggregation pathways that are encountered on directly mixing the components at physiological salt concentrations. The aggregation pathways can also be avoided by using a negatively charged third component as an assembly factor. The use of poly (glutamic acid) as a nucleosome and chromatin assembly factor is described in this chapter. It has been possible to generate spaced nucleosome arrays using histones from all species and cell types that have been examined thus far in our laboratory. However, reaction conditions must be modified in some cases, suggesting that different types of chromatin may have rather different stabilities.
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