ISOLATION OF METAPHASE CHROMOSOMES FROM HELA CELLS |
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Authors: | Joel A. Huberman and Giuseppe Attardi |
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Affiliation: | From the Division of Biology, California Institute of Technology, Pasadena |
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Abstract: | The authors have developed a method for large-scale isolation of metaphase chromosomes from HeLa cells. The distinguishing feature of this method is the use of a pH sufficiently low (about 3) to stabilize the chromosomes against mechanical damage. Many milligrams of fairly pure, morphologically intact chromosomes can be isolated in 8 hr or less of total working time. The isolated chromosomes contain about 2.0 mg of acid-soluble protein, 2.7 mg of acid-insoluble protein and 0.66 mg of RNA for each milligram of DNA. The RNA bound to the isolated chromosomes consists mainly of ribosomal RNA, but there is also a significant amount of 45S RNA. |
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