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Altered electrophoretic behavior of DNA due to short‐time UV‐B irradiation
Author(s) -
Mäueler Winfried,
Kyas Andreas,
Bröcker Frank,
Epplen Jörg T.
Publication year - 1994
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501501214
Subject(s) - ethidium bromide , gel electrophoresis , dna , irradiation , electrophoresis , microbiology and biotechnology , polyacrylamide gel electrophoresis , sodium dodecyl sulfate , chemistry , gel electrophoresis of nucleic acids , staining , biochemistry , biology , enzyme , genetics , physics , nuclear physics
UV‐B irradiation is often inevitable for visualization of DNA fragments after ethidium bromide staining. Three different simple‐repeat‐containing, double‐stranded genomic DNA fragments were analyzed for UV‐B (312 nm) damage using different gel electrophoretic systems. The effects of UV‐B light were obvious after 5 min (31.5 kJ/m 2 ) of irradiation in native polyacrylamide gel electrophoresis (PAGE). Standard single‐strand conformation analyses revealed no alterations while a modification did. Sodium dodecyl sulfate (SDS)‐PAGE was found to be highly sensitive with regard to the detection of damages and their time/dosage dependency. In addition, SDS‐PAGE analysis pointed to different events occurring during UV‐B irradiation. Alterations in DNA conformation were detected in every single strand analyzed after 1 min (6.3 kJ/m 2 ) of UV‐B exposure. Gel retardation analyses revealed significant changes of protein binding to target DNAs after 2 min of irradiation – possibly stemming from structural modifications and/or originating from binding sites for proteins involved in DNA repair.