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Extra resources for Chromatin Structure and Function: Molecular and Cellular Biophysical Methods
Nucleosomal DNA, particularly that containing histone 1, is extremely insoluble in NaCl or magnesium chloride at these concentrations, and therefore precipitates. The resulting precipitate, removed by centrifugation, is known as pellet 2 and the remaining supernatant as supernatant 2. The kinetics of digestion are clear-cut. With sufficient time as much as 80% of the chromatin is solubilized into supernatant 1. By this time, pellet 2 grows to about 50% of the total chromosomal DNA and the supernatant S2, to about 20% of the total chromosomal DNA.
1974) Science 183, 330-332. Ottensmeyer, F. , Hhiting, R. , Schmidt, E. , and Clemens, R. S. (1975) J. Ultrastruct. Res. ~, 193-201. Pooley, A. , Pardon, J. , and Richards, B. M. (1974) J. Mol. BioI. ~, 533-549. Woodcock, C. L. , Safer, J. , and Stanchfield, J. E. (1976) Exp. Cell Res. 22, 101-110. ATIN R. Stewart Gilmour Beatson Institute for Cancer Research Wolfson Laboratory for Molecular Fathology Garscube Estate, Switchback Road, Bearsden Glase;ow, G6l JBD, Scotland WHAT IS CHROMATIN? Most work which has been done with chromatin refers to the dispersed chromosomes of interphase cells.
This is not seen, however, in frozen nuclease digest fragments, especially monomers (VI)' dimers (V2) and trimers (V3). We usually apply the samples to the grids the same day that live cells are harvested. The forces of air-water or air-buffer interphase spreading seem to cause similar gelatinous fibrillar networks and this method is, therefore, avoided in our laboratory. 20 Hany strange structures can be found on an electron microscope grid. Uhich are artifactual and which reflect the structure of chromatin?