A. Asai et al., RESTRICTION-ENDONUCLEASE RECOGNITION AND SOUTHERN HYBRIDIZATION OF BROMODEOXYURIDINE-SUBSTITUTED GENOMIC DNA, Cell proliferation, 26(3), 1993, pp. 271-280
Human glioma cell lines exposed to various concentrations of bromodeox
yuridine (BrdUrd) were studied to determine the effect of BrdUrd subst
itution on restriction endonuclease recognition and Southern hybridiza
tion of genomic DNA. BrdUrd substitution had no effect on the recognit
ion of restriction endonucleases. When the exposure to BrdUrd was 2 h
or less and the BrdUrd substitution rate was less than 40%, there was
no difference in the density of hybridized bands after Southern hybrid
ization using human non-recombinant complementary DNA as a probe. Hybr
idization was suppressed significantly by exposures longer than 24 h o
r BrdUrd substitution rates greater than 40%. These results suggest th
at the BrdUrd substitution rate and the exposure time to BrdUrd influe
nce 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.