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J. Biol. Chem., Vol. 259, Issue 20, 12456-12462, 10, 1984
SM Cohn and MW Lieberman
We have developed an immunological method for isolation and identification
of DNA sequences containing 5-bromo-2'-deoxyuridine (BrdUrd) incorporated
during UV-induced excision-repair synthesis. DNA fragments containing
BrdUrd incorporated during repair synthesis were incubated with goat
anti-BrdUrd and rabbit anti-goat IgG, and the antibody-DNA complexes were
separated from bulk DNA by nitrocellulose filter binding. With this method,
80% of DNA sequences containing BrdUrd-labeled excision-repair sites were
recovered, contaminated with less than 1% of DNA fragments devoid of
excision-repair sites. Recovery of DNA fragments containing repair sites
was independent of size from 2 to 20 kilobases. We have used this method in
conjunction with blot hybridization to demonstrate that repair synthesis
occurs in human ribosomal gene sequences in cells treated with UV.
The use of antibodies to 5-bromo-2'-deoxyuridine for the isolation of DNA sequences containing excision-repair sites
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