Isolation of chromosome clusters from metaphase-arrested HeLa cells
dc.contributor.author | Paulson, James R. | en_US |
dc.date.accessioned | 2006-09-11T18:53:12Z | |
dc.date.available | 2006-09-11T18:53:12Z | |
dc.date.issued | 1982-08 | en_US |
dc.identifier.citation | Paulson, James R.; (1982). "Isolation of chromosome clusters from metaphase-arrested HeLa cells." Chromosoma 85(4): 571-581. <http://hdl.handle.net/2027.42/47359> | en_US |
dc.identifier.issn | 1432-0886 | en_US |
dc.identifier.issn | 0009-5915 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/47359 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=7128277&dopt=citation | en_US |
dc.description.abstract | We have developed a simplified approach for the isolation of metaphase chromosomes from HeLa cells. In this method, all the chromosomes from a cell remain together in a bundle which we call a “metaphase chromosome cluster”. Cells are arrested to 90–95% in metaphase, collected by centrifugation, extracted with non-ionic detergent in a low ionic strength buffer at neutral pH, and homogenised to strip away the cytoskeleton. The chromosome clusters which are released can then be isolated in a crude state by pelleting or they can be purified away from nearly all the interphase nuclei and cytoplasmic debris by banding in a Percoll TM density gradient. — This procedure has the advantages that it is quick and easy, metaphase chromatin is recovered in high yield, and Ca ++ is not needed to stabilise the chromosomes. Although the method does not yield individual chromosomes, it is nevertheless very useful for both structural and biochemical studies of mitotic chromatin. The chromosome clusters also make possible biochemical and structural studies of what holds the different chromosomes together. Such information could be useful in improving chromosome isolation procedures and for understanding suprachromosomal organisation of the nucleus. | en_US |
dc.format.extent | 1444830 bytes | |
dc.format.extent | 3115 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Springer-Verlag | en_US |
dc.subject.other | Developmental Biology | en_US |
dc.subject.other | Eukaryotic Microbiology | en_US |
dc.subject.other | Animal Genetics and Genomics | en_US |
dc.subject.other | Cell Biology | en_US |
dc.subject.other | Biochemistry, General | en_US |
dc.subject.other | Human Genetics | en_US |
dc.subject.other | Life Sciences | en_US |
dc.title | Isolation of chromosome clusters from metaphase-arrested HeLa cells | en_US |
dc.type | Article | en_US |
dc.subject.hlbsecondlevel | Natural Resources and Environment | en_US |
dc.subject.hlbsecondlevel | Molecular, Cellular and Developmental Biology | en_US |
dc.subject.hlbsecondlevel | Ecology and Evolutionary Biology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Laboratory of Molecular Biology, The MRC Centre, Hills Road, CB2 2QH, Cambridge, England; Biophysics Research Division, Institute of Science and Technology, University of Michigan, 48109, Ann Arbor, Michigan, USA | en_US |
dc.contributor.affiliationumcampus | Ann Arbor | en_US |
dc.identifier.pmid | 7128277 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/47359/1/412_2004_Article_BF00327351.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1007/BF00327351 | en_US |
dc.identifier.source | Chromosoma | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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