Restriction endonuclease recognition and southern hybridization of bromodeoxyuridine-substituted genomic DNA.
Human glioma cell lines exposed to various concentrations of bromodeoxyuridine (BrdUrd) were studied to determine the effect of BrdUrd substitution on restriction endonuclease recognition and Southern hybridization of genomic DNA. BrdUrd substitution had no effect on the recognition of restriction endonucleases. When the exposure to BrdUrd was 2h or less and the BrdUrd substitution rate was less than 40%, there was no difference in the density of hybridized bands after Southern hybridization using human non-recombinant complementary DNA as a probe. Hybridization was suppressed significantly by exposures longer than 24 h or BrdUrd substitution rates greater than 40%. These results suggest that the BrdUrd substitution rate and the exposure time to BrdUrd influence the hybridization reaction by a DNA probe. Brief exposure (up to 2 h) to BrdUrd does not influence restriction endonuclease recognition or Southern hybridization of genomic DNA.
Department of Neurological Surgery, University of California, San Francisco.
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A
Asai
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H
Hirai
W J
WJ
Bodell
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T
Hoshino
D001483:Base Sequence; D015139:Blotting, Southern; D001973:Bromodeoxyuridine / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry*; D004262:DNA Restriction Enzymes; D016678:Genome; D005910:Glioma / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000187:drug effects
D004247:DNA; D004262:DNA Restriction Enzymes; D001973:Bromodeoxyuridine