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Refined characterisation of chromosome aberrations in tumours by multicolour banding and electronic mapping resources

Gisselsson Nord, David LU (2001) In Methods in cell science: an official journal of the Society for In Vitro Biology 23(1-3). p.23-28
Abstract
Acquired chromosome abnormalities in tumours often reflect pathogenetic events at the gene level. Multicolour fluorescence in situ hybridisation (FISH) with single-copy probes offers extensive possibilities to characterise chromosome breakpoints in relation to the physical map of the human genome. This approach is based on the construction of comprehensive EST- based maps, combinatorial labelling of probes, and tumour cell preparations optimised for metaphase FISH. Information from several electronically available databases is combined into an integrated physical map, to which clones carrying yeast and bacterial artificial chromosomes are anchored. Extracted DNA or PCR products from these clones are then fluorescently labelled by one or... (More)
Acquired chromosome abnormalities in tumours often reflect pathogenetic events at the gene level. Multicolour fluorescence in situ hybridisation (FISH) with single-copy probes offers extensive possibilities to characterise chromosome breakpoints in relation to the physical map of the human genome. This approach is based on the construction of comprehensive EST- based maps, combinatorial labelling of probes, and tumour cell preparations optimised for metaphase FISH. Information from several electronically available databases is combined into an integrated physical map, to which clones carrying yeast and bacterial artificial chromosomes are anchored. Extracted DNA or PCR products from these clones are then fluorescently labelled by one or several fluors, allowing simultaneous FISH detection of multiple loci. To improve hybridisation efficiency and reduce background fluorescence, standard methods for chromosome preparation from cultured tumour cells are complemented with a prolonged trypsin treatment to obtain complete disaggregation of cells, and exposure of the metaphase spreads to detergent and saline at high temperature, followed by pepsin digestion to remove extracellular matrix and cytoplasmic debris. The resulting colour-banding allows the characterisation of chromosome abnormalities in relation to expressed sequences, even in tumours exhibiting highly complex rearrangements. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Breakpoints, Chromosome banding, Fluorescence in situ hybridisation, Multicolour FISH, Physical maps
in
Methods in cell science: an official journal of the Society for In Vitro Biology
volume
23
issue
1-3
pages
23 - 28
publisher
Springer
external identifiers
  • pmid:11741141
  • scopus:0035707746
ISSN
1381-5741
DOI
10.1023/A:1013129212367
language
English
LU publication?
yes
id
67801982-f1a9-4bb5-81c7-a023c068f05b (old id 1121766)
date added to LUP
2008-06-27 14:51:59
date last changed
2018-05-29 09:37:39
@article{67801982-f1a9-4bb5-81c7-a023c068f05b,
  abstract     = {Acquired chromosome abnormalities in tumours often reflect pathogenetic events at the gene level. Multicolour fluorescence in situ hybridisation (FISH) with single-copy probes offers extensive possibilities to characterise chromosome breakpoints in relation to the physical map of the human genome. This approach is based on the construction of comprehensive EST- based maps, combinatorial labelling of probes, and tumour cell preparations optimised for metaphase FISH. Information from several electronically available databases is combined into an integrated physical map, to which clones carrying yeast and bacterial artificial chromosomes are anchored. Extracted DNA or PCR products from these clones are then fluorescently labelled by one or several fluors, allowing simultaneous FISH detection of multiple loci. To improve hybridisation efficiency and reduce background fluorescence, standard methods for chromosome preparation from cultured tumour cells are complemented with a prolonged trypsin treatment to obtain complete disaggregation of cells, and exposure of the metaphase spreads to detergent and saline at high temperature, followed by pepsin digestion to remove extracellular matrix and cytoplasmic debris. The resulting colour-banding allows the characterisation of chromosome abnormalities in relation to expressed sequences, even in tumours exhibiting highly complex rearrangements.},
  author       = {Gisselsson Nord, David},
  issn         = {1381-5741},
  keyword      = {Breakpoints,Chromosome banding,Fluorescence in situ hybridisation,Multicolour FISH,Physical maps},
  language     = {eng},
  number       = {1-3},
  pages        = {23--28},
  publisher    = {Springer},
  series       = {Methods in cell science: an official journal of the Society for In Vitro Biology},
  title        = {Refined characterisation of chromosome aberrations in tumours by multicolour banding and electronic mapping resources},
  url          = {http://dx.doi.org/10.1023/A:1013129212367},
  volume       = {23},
  year         = {2001},
}