Marine Chemistry, 37 (1992) 239-255 239
Elsevier Science Publishers B.V., Amsterdam
Determination of dissolved nucleic acids in
seawater by the fluorescence dye, ethidium
bromide
Seiji Sakano and Akiyoshi Kamatani 1
Tokyo Universityof Fisheries, Konan 4, Minato-ku, Tokyo 108, Japan
(Received 7 February 1991; revision accepted 30 August 1991 )
ABSTRACT
Sakano, S. and Kamatani, A., 1992. Determination of dissolved nucleic acids in seawater by the flu-
orescence dye, ethidium bromide. Mar. Chem., 37: 239-255.
A new method for the determination of dissolved double-stranded deoxyribonucleic acid (DNA)
and ribonucleicacid (RNA) in seawater was developed, evaluated and used to study the fates of these
nucleic acids in marine ecosystems. These nucleic acids, which were pre-concentrated on a hydroxy-
apatite column, were determined fluorometrically by the use of ethidium bromide dye, which binds
specifically to the double-stranded polynucleotide. No dissolved organic matter coexisting in the pre-
concentrated sample solution interfered in the analysis of DNA and RNA. Column recoveries of DNA
and RNA in a sample volume of up to 11 were 93% and 97%, respectively, and 90% of both at 51. The
detection limits of DNA and RNA concentrated from a 5 1sample by this fluorometric method were
0.6 and 1.1/tg 1-1, respectively. The concentration of dissolved nucleic acids in the waters from Tokyo
Bay and Sagami Bay showed great variation in space and time. DNA ranged from 1 to 32 #g 1-1, and
RNA from below the detection limit to 34/tg 1- I. The total amount of phosphorus in nucleic acids
was an important fraction (12.9 + 8.2%) of the dissolved organic phosphorus (DOP) and showed a
good correlation with DOP.
INTRODUCTION
Phosphorus is an essential element in the structure of nucleic acids, phos-
pholipids and ATP (adenosine triphosphate), and it has an active role in many
life processes such as protein synthesis, genetic coding, photosynthesis, nitro-
gen fixation and innumerable other metabolic pathways. Most of the organic
phosphorus compounds in living tissues are readily excreted and mineralized,
but a portion is metabolized and eliminated as extracellular materials; this
fraction remains in seawater as dissolved organic phosphorus (DOP). The
DOP content varies from below the detection limit (0.5 #g 1-1 ) to 52/tg 1-1;
IAuthor to whom correspondence should be addressed.
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