Improvements in the fluorimetric determination of the RNA and DNA content of individual marine fish larvae.

Clemmesen, Catriona (1993) Improvements in the fluorimetric determination of the RNA and DNA content of individual marine fish larvae. Open Access Marine Ecology Progress Series, 100 . pp. 177-183.

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Abstract

The RNA/DNA ratio is a useful indicator of the nutritional condition of fish larvae. The presented analytical procedure is an improvement of Clemmesen's (Meeresforschung 32: 134-143,
1988) methodology which involves purification of fish larvae tissue homogenates and subsequent
fluorescence-photometric measurements using specific nucleic acid dyes. The modifications concern the homogenization and nucleic acid extraction procedures. A 'shaking mill' was compared to a potter Elvehjem microhomogenizer and a reduction in the washing and purification steps was achieved.
Treatment of samples with ribonuclease A and subsequent fluorescence measurement using ethidium bromide was given preference compared to the DNA-bisbenzimidazole determinations due to problems arising from high self-fluorescence of the samples and the influence of 'quenching' substances disturbing the DNA-bisbenzmidazole determinations. Different RNase concentrations and their influences on RNA and DNA were checked. Recovery rates of standard RNA and DNA 'spikes' were determined. Fish larvae samples were analysed with the previous and the improved modified procedure and a correction factor to compare results measured with the 2 procedures was calculated.
With the presented method the physiological condition of individual larvae and the amount of variability can be determined.

Document Type: Article
Research affiliation: OceanRep > GEOMAR > FB3 Marine Ecology > FB3-EV Marine Evolutionary Ecology
Refereed: Yes
Open Access Journal?: No
ISSN: 0171-8630
Date Deposited: 17 Nov 2009 10:03
Last Modified: 07 Mar 2018 13:31
URI: http://oceanrep.geomar.de/id/eprint/6759

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