Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes

Heinz Ulrich G Weier, Daniel Polikoff, John J. Fawcett, Karin M. Greulich, Kwang Ho Lee, Scott Cram, Verne M. Chapman, Joe Gray

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Mouse metaphase chromosomes were purified by flow sorting from the murine fibroblast cell line Mus spretus clone 5A. We sorted chromosomes that fell into five individual peaks based on the Hoechst 33258/chromomycin A3 DNA histogram: three peaks corresponding to the least amount of DNA and two peaks representing chromosomes with the most DNA content. This is the first example of the successful application of bivariate flow karyotyping to murine chromosome sorting. We then applied primer-directed in vitro DNA amplification using the polymerase chain reaction (PCR) to generate and label larger amounts of chromosome-specific DNA. In situ hybridization showed specific binding of the PCR products to mouse chromosomes Y, 19, 18, 3, and X as well as chromosomes 1 and 2. The combination of chromosome sorting from the M. spretus cell line and PCR proved to be highly valuable for generation of pools of DNA fragments that exhibit specific binding to mouse chromosomes and can be used to identify and delineate mouse metaphase chromosomes.

Original languageEnglish (US)
Pages (from-to)641-644
Number of pages4
JournalGenomics
Volume21
Issue number3
DOIs
StatePublished - Jun 1994
Externally publishedYes

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Paintings
Chromosomes
DNA
Metaphase
Polymerase Chain Reaction
Chromomycin A3
Bisbenzimidazole
Chromosomes, Human, Pair 19
Cell Line
Karyotyping
Chromosomes, Human, Pair 2
Chromosomes, Human, Pair 1
In Situ Hybridization
Clone Cells
Fibroblasts

ASJC Scopus subject areas

  • Genetics

Cite this

Weier, H. U. G., Polikoff, D., Fawcett, J. J., Greulich, K. M., Lee, K. H., Cram, S., ... Gray, J. (1994). Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes. Genomics, 21(3), 641-644. https://doi.org/10.1006/geno.1994.1326

Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes. / Weier, Heinz Ulrich G; Polikoff, Daniel; Fawcett, John J.; Greulich, Karin M.; Lee, Kwang Ho; Cram, Scott; Chapman, Verne M.; Gray, Joe.

In: Genomics, Vol. 21, No. 3, 06.1994, p. 641-644.

Research output: Contribution to journalArticle

Weier, HUG, Polikoff, D, Fawcett, JJ, Greulich, KM, Lee, KH, Cram, S, Chapman, VM & Gray, J 1994, 'Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes', Genomics, vol. 21, no. 3, pp. 641-644. https://doi.org/10.1006/geno.1994.1326
Weier HUG, Polikoff D, Fawcett JJ, Greulich KM, Lee KH, Cram S et al. Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes. Genomics. 1994 Jun;21(3):641-644. https://doi.org/10.1006/geno.1994.1326
Weier, Heinz Ulrich G ; Polikoff, Daniel ; Fawcett, John J. ; Greulich, Karin M. ; Lee, Kwang Ho ; Cram, Scott ; Chapman, Verne M. ; Gray, Joe. / Generation of Five High-Complexity Painting Probe Libraries from Flow-Sorted Mouse Chromosomes. In: Genomics. 1994 ; Vol. 21, No. 3. pp. 641-644.
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