Preparation of metaphase chromatin of without the loss of repressed RNA synthesis |
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Authors: | Fumio Sawada Yasuhiko Miyauchi Hiroshi Tanaka Shinji Matsumoto |
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Affiliation: | Division of Chemistry, National Institute of Radiological Sciences, 9-1, Anagawa-4-chome, Chiba-shi 260, Japan |
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Abstract: | A novel method for the preparation of intact chromatin from the slime mold which retains the property of RNA synthesis is described. Preparations from G2-cells were highly active, while those from metaphase-cells were inactive. The plasmodial cells were disrupted by gentle homogenization on a polyethylene sieve in a neutral isotonic sucrose medium containing Mg++, deoxycholate and EGTA, a Ca++-chelating agent. The nuclei were lysed in a hypotonic buffer without use of EDTA and chromatin was precipitated by centrifugation after addition of Mg++. |
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Keywords: | DOC potassium deoxycholate DTT dithiothreitol EGTA TCA trichloroacetic acid |
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