Isolation of a nuclear DNA synthesizing complex from human sperm
Education, Medical, Undergraduate
A simplified procedure is described for isolating a human sperm nuclear DNA-synthesizing complex which does not involve the use of detergent or necessitate the prior purification of membrane-free nuclei. Selective disruption of sperm heads is accomplished by the use of dithiothreitol, trypsin, and deoxyribonuclease. The particulate complex is freed of soluble components by sedimentation through a solution of 19% (w/v) Metrizamide and further purified by buoyant density centrifugation. The characteristics of this endogenously templated nuclear DNA polymerase differ from other sperm-associated polymerases that have been reported. © 1977 Academic Press, Inc. All rights of reproduction in any form reserved.
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