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Preparation of metaphase chromatin of Physarumpolycephalum without the loss of repressed RNA synthesis
Authors:Fumio Sawada  Yasuhiko Miyauchi  Hiroshi Tanaka  Shinji Matsumoto
Institution:Division of Chemistry, National Institute of Radiological Sciences, 9-1, Anagawa-4-chome, Chiba-shi 260, Japan
Abstract:A novel method for the preparation of intact chromatin from the slime mold Physarumpolycephalum> which retains the invivo 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++.
Keywords:DOC  potassium deoxycholate  DTT  dithiothreitol  EGTA  TCA  trichloroacetic acid
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