Hoechst Fluorometric quantitation of DNA

Tuesday, November 18, 2003

Description
The concentration of DNA in solutions containing contaminating protein or RNA can be determined quite accurately by measuring the increased fluorescence of the Hoechst dye 33258 upon intercalating into dsDNA.

Procedure
1) Samples to be measured should be diluted to 1ml in H2O or TE, pH8.0.

2) 1ml of Hoechst buffer (4M NaCl, 100mM NaP04, pH7.4, 0.2ug/ml Hoechst 33258) is added to each sample.

3) Measure fluorescence at an emission wavelength of 458nM and an excitation wavelength of 356nM.

DNA concentrations can be measured by comparison of the emission intensities of the unknown samples with those generated by a series of samples of known DNA concentrations.

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Hoechst Fluorometric quantitation of DNA