Isolation of metaphase chromosomes from HeLa cells.
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.
D002455:Cell Division; D002467:Cell Nucleus / Q000032:analysis; D002875:Chromosomes / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D009363:Neoplasm Proteins / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis
D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm