Quantification of total RNA by ethidium bromide fluorescence may not accurately reflect the RNA mass

Journal of Biochemical and Biophysical Methods
L Kerkhof

Abstract

The fluorescent signal from purified large and small ribosomal RNA subunits stained with ethidium bromide was quantified using agarose gels and image analysis. A significantly smaller fluorescent signal was observed for 23 S rRNA compared to 16 S rRNA when normalized to mass. Therefore, small changes in amounts of 23 S or 16 S rRNA within a sample can have a profound impact on observed bulk fluorescence. A comparison of 23 S/16 S rRNA fluorescence ratios for 4 marine bacteria grown in batch culture indicates that the lower fluorescence of 23 S rRNA is widespread. The 23 S/16 S rRNA fluorescence ratios also suggest that intracellular concentrations of ribosomal subunits do not always adhere to a stoichiometry of 2.0.

References

Oct 1, 1966·Analytical Biochemistry·J B Le Pecq, C Paoletti
Jul 18, 1993·Biochimica Et Biophysica Acta·A BonincontroG Risuleo
Nov 1, 1957·Proceedings of the Society for Experimental Biology and Medicine·A F CARLUCCI, D PRAMER

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