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Open Access Green as soon as Postprint is submitted to ZB.
Rate of DNA synthesis determined by flow cytometry using the BrdUrd/Hoechst technique in combination with propidium-iodide staining.
Exp. Cell Res. 139, 111-115 (1982)
Cells were exposed to BrdUrd for intervals of between 0 and 3 h, stained with a mixture of 33258 Hoechst and propidium iodide (PI) and analysed in a two-parametrical flow cytometer. The BrdUrd-quenched Hoechst fluorescence gives information about where the cells were located at time 0 of the incubation, and the DNA-specific PI fluorescence gives information about the actual location of the cells in the cycle. Narrow windows in the Hoechst fluorescence were selected. The location of the corresponding cells in the S phase, as determined from the peaks of the projection on the PI-axis, yielded information about how far the cells had travelled through S phase during BrdUrd incubation. Thereby, the DNA synthesis rate in subcompartments of S phase was determined and showed maximum values at approximately mid-S. In addition, information is obtained about the whole duration of the S phase and the spread of transit velocities through subcompartments of S.
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Publication type
Article: Journal article
Document type
Scientific Article
ISSN (print) / ISBN
1090-2422
e-ISSN
0014-4827
Journal
Experimental Cell Research
Quellenangaben
Volume: 139,
Issue: 1,
Pages: 111-115
Publisher
Academic Press
Publishing Place
Orlando, Fla.
Reviewing status
Peer reviewed
Institute(s)
Institut für Experimentelle Hämatologie