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PMID: 6166650 Published · ppublish English Journal Article

Fluorometric determination of DNA and RNA in Chlamydomonas using ethidium bromide.

Journal of biochemical and biophysical methods ·Vol. 4 ·No. 5-6 ·1981-06-00 ·Pages 271-7

Valle O, Lien T, Knutsen G

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.

MeSH Terms
Chlamydomonas/analysis DNA/analysis Deoxyribonucleases Ethidium Kinetics RNA/analysis Ribonucleases Spectrometry, Fluorescence/methods
Chemicals
RNA DNA Deoxyribonucleases Ribonucleases Ethidium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Valle O
Lien T
Knutsen G
Article Info
Journal
Journal of biochemical and biophysical methods
Abbr.
J Biochem Biophys Methods
ISSN
0165-022X
Published
1981-06-00
Pages
271-7
Language
English
Region
Netherlands
NLM ID
7907378
Subset
IM
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