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A microfluorometric method for quantifying RNA and DNA in terrestrial insects

Author(s): M. Kyle | T. Watts | J. Schade | J.J. Elser

Journal: Journal of Insect Science
ISSN 1536-2442

Volume: 3;
Start page: 1;
Date: 2003;
Original page

Keywords: Ribogreen | Drosophila sp. | Curculionidae | Growth Rate Hypothesis | microplate fluorometry | N-lauroylsarcosine | nucleic acids

Evidence is accumulating for a mechanistic linkage between body phosphorus content and growth and reproduction of individual organisms, due in part to variation in allocation of resources to ribosomal RNA. Testing this connection requires reliable methods of quantifying the nucleic acid content of individual organisms. Although methods for quantifying nucleic acids are available for a wide array of organisms, adaptation of such methods for study of insects has been neglected. Sensitive stains and high throughput fluorometric measurements are now available that substantially improve past methodologies. Here we present methods for the extraction and quantification of insect RNA and DNA based on the use of N-lauroylsarcosine and sonication for extraction, the nucleases RNase and DNase, and the use of microplate fluorescent assays to quantify nucleic acids as percent of body weight in insects. We illustrate the method using Drosophila and curculionid weevils.
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