Molecular characterization of flow-sorted mammalian centromeres

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This is the final report of a three-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The project involved experiments directed towards developing a molecular characterization of the centromere region of mammalian chromosomes. Attempts to purify this essential chromosomal locus by conventional methods have thus far been unsuccessful. However, preliminary data obtained in collaboration with the National Flow Cytometry Resource (NFCR) showed that it is possible to purify a chromosome fragment that is present in certain cultured mouse cell lines and has all the properties expected of an intact centromere region. To begin sorting ... continued below

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9 p.

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Hamkalo, B.A.; Henschen, A. & Parseghian, M.H. December 31, 1998.

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Description

This is the final report of a three-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The project involved experiments directed towards developing a molecular characterization of the centromere region of mammalian chromosomes. Attempts to purify this essential chromosomal locus by conventional methods have thus far been unsuccessful. However, preliminary data obtained in collaboration with the National Flow Cytometry Resource (NFCR) showed that it is possible to purify a chromosome fragment that is present in certain cultured mouse cell lines and has all the properties expected of an intact centromere region. To begin sorting this minichromosome for the identification of proteins preferentially associated with centromere regions, standard buffers utilized in chromosome sorting were evaluated for potential effects on maintenance of chromosomal proteins during sorting. The data indicate that the presence of several buffer constituents results in the extraction of all but a few chromosomal proteins. The subsequent use of a magnesium sulfate buffer resulted in the sorting of mouse chromosomes that do not suffer a significant loss of proteins. Several DNA stains were also evaluated for causing protein dissociation, but no significant losses were observed. Although flow-sorted chromosomes have been used extensively for DNA analysis and cloning, this is a pioneering effort by the NFCR, and its collaborators, to exploit chromosome sorting capabilities for the analysis of chromosomal proteins.

Physical Description

9 p.

Notes

OSTI as DE98001516

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  • Other Information: PBD: [1998]

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  • Other: DE98001516
  • Report No.: LA-UR--97-3238
  • Grant Number: W-7405-ENG-36
  • DOI: 10.2172/560745 | External Link
  • Office of Scientific & Technical Information Report Number: 560745
  • Archival Resource Key: ark:/67531/metadc697627

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Office of Scientific & Technical Information Technical Reports

Reports, articles and other documents harvested from the Office of Scientific and Technical Information.

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  • December 31, 1998

Added to The UNT Digital Library

  • Aug. 14, 2015, 8:43 a.m.

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  • May 20, 2016, 4:28 p.m.

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Hamkalo, B.A.; Henschen, A. & Parseghian, M.H. Molecular characterization of flow-sorted mammalian centromeres, report, December 31, 1998; New Mexico. (digital.library.unt.edu/ark:/67531/metadc697627/: accessed October 16, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.