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This solution should be used very cold (not warmer than -10 C), because the lower temperatures help to increase its viscosity. 2 M sucrose and only 5% glycerol. 5 to 2 g/10 ml). PMSF (phenyl-methyl-sulfonyl-fluoride) is a Serine-protease inhibitor. It is soluble in isopropanol at room temperature. For storage, the solution is kept at -20 C, where the PMSF crystallizes. It can be redissolved by a brief heating at 57 C, and kept at room temperature until needed; if kept on ice, it will re-crystallize!

Nuclei are contaminated by red blood cells and other debris after the first centrifugation, due to an excessive amount of tissue being processed at one time. D. Nuclei are contaminated by cytoplasmic tags due to insufficient homogenization. III Experimental 39 homogenization buffer containing an additional 10% of glycerol. Usually, this step is carried out in about 50 ml (two SW27 tubes). The nuclei are resuspended by bubbling and scraping the side of the tube with a plastic pipet (avoid glass pipets, as nuclei stick much more to them), then, carefully aspirate up and down several times, making sure not to produce foaming.

Stir in warm room (57 C) until dissolved, then add the other components. Store at _20DC at least overnight. This solution should be used very cold (not warmer than -10 C), because the lower temperatures help to increase its viscosity. 2 M sucrose and only 5% glycerol. 5 to 2 g/10 ml). PMSF (phenyl-methyl-sulfonyl-fluoride) is a Serine-protease inhibitor. It is soluble in isopropanol at room temperature. For storage, the solution is kept at -20 C, where the PMSF crystallizes. It can be redissolved by a brief heating at 57 C, and kept at room temperature until needed; if kept on ice, it will re-crystallize!

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