Fluorometric determination of DNA and RNA in Chlamydomonas using ethidium bromide

Journal of Biochemical and Biophysical Methods
O ValleG Knutsen

Abstract

By using the fluorescence enhancement of ethidium bromide bound to nuclei acid, a very rapid, simple and sensitive assay of DNA in the green alga Chlamydomonas has been devised. Total fluorescence (DNA + RNA) was determined by complex formation with ethidium bromide in a cell lysate made by mixing cell samples with lauroyl sarcosinate, EDTA and NaOH and incubating the mixture for 5 min at room temperature followed by neutralization. For determination of DNA the RNA was digested by incubating the cell sample in te alkaline lysis solution for 45 min at 60 degrees C followed by neutralization, and complex formation with ethidium bromide. Quenching of the fluorescence due to cellular pigments was corrected for using an internal DNA standard.

References

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Oct 1, 1966·Analytical Biochemistry·J B Le Pecq, C Paoletti

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Citations

Jul 11, 2001·Genes & Development·J G Umen, U W Goodenough
Jul 9, 2004·Applied and Environmental Microbiology·Freddie H SharkeyRoger Marchant
Jan 9, 2009·BMC Systems Biology·Nanette R Boyle, John A Morgan
Mar 20, 2012·Journal of Applied Phycology·Anna M J KliphuisRené H Wijffels
Dec 19, 2009·Expert Opinion on Drug Delivery·Maria Luisa Bondì, Emanuela Fabiola Craparo
May 11, 2007·Expert Opinion on Drug Delivery·Mayank D Bhavsar, Mansoor M Amiji
May 4, 2011·Biotechnology and Bioengineering·Anna M J KliphuisRené H Wijffels

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