Separation of large quantities of chinese hamster chromosomes by velocity sedimentation at unit gravity followed by flow sorting (FACS II) |
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Authors: | JG Collard A Tulp J Stegeman J Boezeman FW Bauer JF Jongkind A Verkerk |
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Institution: | 1. The Netherlands Cancer Institute, Divisions of Cell Biology and Biochemistry, 1066 CX Amsterdam, The Netherlands;2. University of Nijmegen, Department of Dermatology, Nijmegen, The Netherlands;3. Erasmus University, Department of Cell Biology, Rotterdam, The Netherlands |
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Abstract: | Separation of large quantities of isolated metaphase chromosomes of Chinese hamster cells was performed by velocity sedimentation at unit gravity in a specially designed sedimentation chamber. This simple and easy technique results in chromosome fractions of relatively high purity as determined by flow cytometry and microscopy. Up to 1010 chromosomes can be processed depending upon the size of the sedimentation device, and enrichments up to 10 times of individual chromosomes were achieved. In addition, further chromosome purification was performed by fluorescence activated flow sorting using fractions, pre-enriched at unit gravity. The flow sorted chromosomal fractions were pure according to flow cytometric analyses. The combination of l g sedimentation and flow-sorting opens the possibility for preparative chromosome sorting by reducing the flow sorting time considerably. |
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Keywords: | Address for offprint requests: The Netherlands Cancer Institute Department of Experimental Cytology Division of Cell Biology 121 Plesmanlaan 1066 CX Amsterdam The Netherlands |
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