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A microfluorometric assay has been developed for the quantitation of de novo DNA synthesis. The assay reliably detects newly synthesized DNA in individual cells and provides a relative measure of the proportion of new DNA in each cell. When combined with microscopic techniques for cell identification, selected subpopulations can be examined in samples containing a complex mixture of interacting cell types. The assay employs the quenching of Hoechst-33258 fluorescence by bromodeoxyuridine incorporated into newly synthesized DNA. The ability of BUdR to quench H-33258 fluorescence is lost after a brief exposure of ultraviolet fluorescence excitation. Therefore, quenched and unquenched fluorescence intensity measurements from the same cell can be compared. Increases in fluorescence intensity occur only in cells that have synthesized DNA after the addition of BUdR. In addition the change in fluorescence intensity is proportional to the degree of BUdR substitution within the range of the assay and provides a relative measure of DNA synthesis.
L K Gordon, and
C W Parker
March 1986,
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L K Gordon, and
C W Parker
January 1977,
The Italian journal of biochemistry,
L K Gordon, and
C W Parker
October 1983,
Archives of biochemistry and biophysics,
L K Gordon, and
C W Parker
January 2008,
Nucleic acids symposium series (2004),
L K Gordon, and
C W Parker
November 2004,
Nucleic acids research,
L K Gordon, and
C W Parker
April 2003,
Molecular and cellular biology,
L K Gordon, and
C W Parker
October 2003,
Molecular biotechnology,
L K Gordon, and
C W Parker
October 2008,
Nucleic acids research,
L K Gordon, and
C W Parker
August 2018,
Nature biotechnology,
L K Gordon, and
C W Parker
January 2012,
Methods in molecular biology (Clifton, N.J.),
