A filter elution assay for the simultaneous detection of DNA double and single strand breaks

Junxuan Lu, Mark R. Kaeck, Cheng Jiang, Gretchen Garcia, Henry J. Thompson

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

This paper reports a gravity-flow filter elution assay that permits the simultaneous detection of DNA double strand breaks (DSBs, as neutral elutable DNA) and DNA single strand breaks (SSBs, as alkaline elutable DNA). This assay is based on a modification of a conventional gravity-flow alkaline elution procedure. The elution fraction resulting from cell lysis before the application of the alkaline elution buffer provides an estimate of DNA DSBs, while the fraction eluted by the alkaline buffer (pH 12.3) measures the extent of DNA SSBs. Data show that, in addition to its good detection sensitivity for SSBs, this method provides a sensitive measure of DNA DSBs. As an illustration of its specificity, this method unambiguously distinguished DNA SSBs induced by X-irradiation of mouse leukemia L1210 cells from DNA DSBs occurring in cells undergoing nucleosomal DNA fragmentation and apoptosis. The neutral elution fraction is a good predictor of loss of cell viability induced either by selenite at pharmacological levels or by growth factor depletion. The filter elution method reported is more specific and informative than a nick translation assay for the detection of DNA strand breaks.

Original languageEnglish (US)
Pages (from-to)227-233
Number of pages7
JournalAnalytical Biochemistry
Volume235
Issue number2
DOIs
StatePublished - Mar 15 1996

Fingerprint

Single-Stranded DNA Breaks
Double-Stranded DNA Breaks
Assays
DNA
Gravitation
Buffers
Selenious Acid
Leukemia L1210
DNA Breaks
DNA Fragmentation
Cell Survival
Intercellular Signaling Peptides and Proteins
Pharmacology
Apoptosis
1,2-di-(4-sulfamidophenyl)-4-butylpyrazolidine-3,5-dione
Cells
Irradiation

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lu, Junxuan ; Kaeck, Mark R. ; Jiang, Cheng ; Garcia, Gretchen ; Thompson, Henry J. / A filter elution assay for the simultaneous detection of DNA double and single strand breaks. In: Analytical Biochemistry. 1996 ; Vol. 235, No. 2. pp. 227-233.
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A filter elution assay for the simultaneous detection of DNA double and single strand breaks. / Lu, Junxuan; Kaeck, Mark R.; Jiang, Cheng; Garcia, Gretchen; Thompson, Henry J.

In: Analytical Biochemistry, Vol. 235, No. 2, 15.03.1996, p. 227-233.

Research output: Contribution to journalArticle

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