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• W209659648 abstract "Chromosomes are ordinarily obtained from cells during interphase and are, therefore, in the extended form known as chromatin. The advancement in the understanding of chromosomal structure and function has been made possible by the development of new methods for the handling of chromatin and chromosomal constituents. The isolation of chromatin is based upon differential centrifugation followed by sucrose density gradient centrifugation. Chromatin is among the most pelletable components of a tissue homogenate. The tissue is, therefore, ground in a suitable medium, freed of unruptured cells and membrane fragments by filtration, and sedimented at 1000–4000 g, conditions that do not bring down mitochondria. The pellet is then washed by repeated suspension and pelleting, finally layered on sucrose solution, and centrifuged for an appropriate period. By these methods, 60–75% or more of the DNA present in the original tissue is recovered as purified chromatin. The basic steps for the isolation of the highly contracted metaphase chromosomes are (1) accumulation of a large proportion of cells in metaphase by treatment with colchieine or other mitotic poison, (2) homogenization of the cells without damage to the chromosomes, (3) separation of the released chromosomes from cell debris. The separation of chromosomal nucleoprotein into its component entities can now be accomplished by the methods that are relatively mild and nondestructive as compared to those used in the past. An excellent example is the separation of histones from DNA by banding in a cesium chloride density gradient." @default.
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• W209659648 date "1968-01-01" @default.
• W209659648 modified "2023-10-11" @default.
• W209659648 title "[96] Isolation and characterization of chromosomal nucleoproteins" @default.
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• W209659648 doi "https://doi.org/10.1016/0076-6879(67)12116-2" @default.
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