Preparative dual-beam sorting of the human Y chromosome and in situ hybridization of cloned DNA probes.

C. Cremer, G. Rappold, Joe Gray, C. R. Müller, H. H. Ropers

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Bivariate Hoechst/chromomycin flow karyotypes for chromosomes from a Chinese hamster-human hybrid cell line (CH-Y-VII) were established that have retained a human Y chromosome. These bivariate flow karyotypes showed the human Y chromosome to be completely separated from the peaks for the Chinese hamster chromosomes. In preparative dual-beam sorting experiments, 3 X 10(6) chromosomes were sorted from the Y peak into frozen petri dishes. An examination of Q-banded samples of sorted chromosomes revealed that 82% +/- 5% of them were human Y chromosomes. The DNA from the sorted chromosomes (approximately 250 ng) was isolated and used to establish a genomic library (vector lambda gt WES. lambda B). Three clones (YACG 45, 52, 54) of this library containing inserts of repetitive human DNA were used for chromosomal localization by means of in situ hybridization to metaphase spreads of male human lymphocytes and of CH-Y-VII cells. In all three cases, a significant binding to the human Y chromosome was observed. A more detailed study of the chromosomal distribution of sequences homologous to the insert of YACG 45 suggested the existence of minor binding sites on several human autosomes. Southern blot analysis revealed the existence of other human specific sequences without Y chromosome specificity.

Original languageEnglish (US)
Pages (from-to)572-579
Number of pages8
JournalCytometry
Volume5
Issue number6
StatePublished - Nov 1984
Externally publishedYes

Fingerprint

Chromosomes, Human, Y
DNA Probes
In Situ Hybridization
Chromosomes
Cricetulus
Karyotype
Chromomycins
Chromosomes, Human, Pair 10
Chromosomes, Human, Pair 6
Genomic Library
Hybrid Cells
Y Chromosome
DNA
Metaphase
Sequence Homology
Southern Blotting
Libraries
Clone Cells
Binding Sites
Lymphocytes

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Endocrinology
  • Hematology
  • Pathology and Forensic Medicine

Cite this

Cremer, C., Rappold, G., Gray, J., Müller, C. R., & Ropers, H. H. (1984). Preparative dual-beam sorting of the human Y chromosome and in situ hybridization of cloned DNA probes. Cytometry, 5(6), 572-579.

Preparative dual-beam sorting of the human Y chromosome and in situ hybridization of cloned DNA probes. / Cremer, C.; Rappold, G.; Gray, Joe; Müller, C. R.; Ropers, H. H.

In: Cytometry, Vol. 5, No. 6, 11.1984, p. 572-579.

Research output: Contribution to journalArticle

Cremer, C, Rappold, G, Gray, J, Müller, CR & Ropers, HH 1984, 'Preparative dual-beam sorting of the human Y chromosome and in situ hybridization of cloned DNA probes.', Cytometry, vol. 5, no. 6, pp. 572-579.
Cremer, C. ; Rappold, G. ; Gray, Joe ; Müller, C. R. ; Ropers, H. H. / Preparative dual-beam sorting of the human Y chromosome and in situ hybridization of cloned DNA probes. In: Cytometry. 1984 ; Vol. 5, No. 6. pp. 572-579.
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