Isopycnic centrifugation of mammalian metaphase chromosomes in Nycodenz.
Separation of chromosomes on the basis of buoyant density has proved difficult due to their high density and sensitivity to high ionic strength. Nycodenz is a new non-ionic gradient medium which offers significant advantages over previously described media for separation of chromosomes. The purpose of this study was to examine the banding characteristics of isolated mammalian metaphase chromosomes in Nycodenz gradients. The results indicate that chromosomes can be efficiently concentrated and separated from nuclei using Nycodenz gradients. Furthermore, chromosomes density-labeled with bromodeoxyuridine (BrdU) can be separated from unlabeled chromosomes in Nycodenz gradients. Nycodenz does not appear to alter chromosome morphology or protein complement. The introduction of Nycodenz represents a significant new tool for use in chromosome separation and purification.
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods*; D002460:Cell Line; D002467:Cell Nucleus; D002499:Centrifugation, Density Gradient; D002500:Centrifugation, Isopycnic; D002875:Chromosomes / Q000032:analysis / Q000187:drug effects / Q000648:ultrastructure; D006224:Cricetinae; D005260:Female; D007463:Iodobenzoates / Q000494:pharmacology; D007472:Iohexol; D008677:Metaphase; D008793:Metrizamide / Q000494:pharmacology; D009698:Nucleoproteins / Q000032:analysis; D010053:Ovary; D014283:Triiodobenzoic Acids / Q000494:pharmacology
D007463:Iodobenzoates; D009698:Nucleoproteins; D014283:Triiodobenzoic Acids; D007472:Iohexol; D001973:Bromodeoxyuridine; D008793:Metrizamide