Comparing RNA:DNA ratio methods in a study relating recent growth rates in juvenile flatfish with coastal habitat conditions

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

An ecosystem monitoring program was performed in the context of studying impacts of coastal nourishments in Dutch coastal waters. One of the research questions was determining variation in growth rate of juvenile flatfish in various habitats of the Dutch coast. These shallow areas are considered nurseries for a number of flatfish species, as they provide shelter and food. To get a better understanding of the habitat quality of these nursery areas and possible influence of the nourishments on them, juvenile sole -Solea solea-, plaice -Pleuronectes platessa- and brill - Scophthalmus rhombus- were captured along a gradient of biological, chemical and physical factors. On board these fish were frozen, and in the lab the white muscle tissue was used for the RNA:DNA ratio determination. The amount of RNA and DNA was quantified using two fluorescent methods, the first using the commonly used but highly toxic ethidium bromide -EB- and the second using a new method with less toxic nucleic acid specific dyes. The staining of the nucleic acids was performed in a crude lysate, followed by a fluorescence measurement. Our results indicate that the methods differ in the absolute amounts of DNA and RNA, most likely caused because EB is a non-specific binding agent. Despite this difference, the methods indicate comparable ratios making both methods useful in studies on recent growth rates. We favour the less toxic method, as EB is no longer allowed in many labs, and used this method to analyse the relation between recent growth and the habitat characteristics depth, food availability, sediment, temperature and salinity.
Original languageEnglish
Title of host publicationAbstract book - 10th International Symposium Flatfish
Pages73-73
Publication statusPublished - 2017
Event10th International Symposium Flatfish - Saint-Malo, France
Duration: 11 Nov 201716 Nov 2017
Conference number: 10th

Conference/symposium

Conference/symposium10th International Symposium Flatfish
Country/TerritoryFrance
CitySaint-Malo
Period11/11/1716/11/17

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