Fluorometric determination of DNA in fixed tissue using ethidium bromide

The measurement of DNA in tissue samples fixed in ethanol/acetic acid is described. Small, fixed tissue samples are digested by warm alkaline treatment followed by neutralization with HCl, and DNA is determined by complex formation with the dye ethidium bromide (EB). When standard DNA from calf thym...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 83; no. 2; pp. 703 - 708
Main Authors Andersen, Knut-Jan, Skagen, Dankert W.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.01.1977
Subjects
Animals
DNA - analysis
Ethidium
Intestine, Small - analysis
Kidney - analysis
Liver - analysis
Rats
Spectrometry, Fluorescence - methods
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Summary:The measurement of DNA in tissue samples fixed in ethanol/acetic acid is described. Small, fixed tissue samples are digested by warm alkaline treatment followed by neutralization with HCl, and DNA is determined by complex formation with the dye ethidium bromide (EB). When standard DNA from calf thymus was treated similarly, a hyperchromicity of 8–12% and a reduction in fluorescence intensity of the EB-DNA complex to 55% was observed. The NaOH concentration (0.5–2.0 mol/liter) or the temperature (50–60°C) used for the digestion of tissue, as well as subsequent ribonuclease or protease treatment had no effect on the observed tissue DNA concentrations.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(77)90075-6