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The World of Protozoa, Rotifera, Nematoda and Oligochaeta

Ref ID : 4137

Reinhard Corr and Wolfgang Eichler; Isolation and Properties of Macronuclei from Tetrahymena thermophila. J.Protozool. 32(1):45-49, 1985

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A simple and efficient method is described for the isolation of macronuclei from Tetrahymena thermophila (7B). The steps involved are deciliation and removal of the mucocysts' contents by dibucaine treatment, digitonin mediated lysis, differential centrifugations, and finally isopycnic sucrose density gradient centrifugation. Judging from the distribution of marker enzymes and electron microscopy, the macronuclei obtained were free of cytoplasmic and particulate contamination and were highly active in endogenous RNA-synthesis (1.5 pmol UTP incorporation/ng DNA-min at 30 degrees C). The ratio of protein:RNA:DNA was 2.0:0.33:1.0 (weight) and each macronucleus contained an average of 17 pg DNA. The average yield of isolation was 50%.