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Fluorometric determination of DNA in fixed tissue using ethidium bromide

✍ Scribed by Knut-Jan Andersen; Dankert W. Skagen

Publisher: Elsevier Science
Year: 1977
Tongue: English
Weight: 359 KB
Volume: 83
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(77)90075-6

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✦ Synopsis

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 S-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 (SO-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. 703

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