Physical basis for detection of DNA double-strand breaks using neutral filter elution

Journal Article · Sat Dec 01 04:00:00 EST 1990 · Radiation Research; (USA)

DOI:https://doi.org/10.2307/3577846· OSTI ID:6209830

Wlodek, D; Olive, P L ^[1]

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1. B.C. Cancer Research Centre, Vancouver (Canada)

Results using neutral filter elution are difficult to explain if this method detects only DNA double-strand breaks (DSBs). In an attempt to understand neutral filter elution, the size of DNA pieces eluted from filters was measured using pulsed-field gel electrophoresis. Contrary to expectation, the size of the pieces was independent of radiation dose and time of elution, and much smaller (approximately 460 kb) than anticipated based on the expected number of DSBs induced. Shearing of the DNA molecule, the presence of nonspecific nucleases, and the influence of DNA-associated proteins were examined but could not explain our results. Consequently, we propose that cell lysis causes swelling of the DNA gel, and the exposed fraction of DNA on the surface of the gel is then sheared as the elution solution flows through the filter. We suggest that the rate of DNA elution measured using neutral filter elution is dependent upon the number of DSBs present, the composition of the eluting solution, especially with regard to the presence of molecules which can influence chromatin swelling on the filter, and the conformation or packaging of DNA before lysis.

• OSTI ID: 6209830
• Journal Information: Radiation Research; (USA), Journal Name: Radiation Research; (USA) Vol. 124:3; ISSN 0033-7587; ISSN RAREA
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
DNA
ELECTROPHORESIS
ENZYMES
ESTERASES
FILTRATION
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
HYDROLASES
IN VITRO
LYSIS
NUCLEASES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
RADIATION EFFECTS
RADIOINDUCTION
SEPARATION PROCESSES
STRAND BREAKS