FLUORESCENT MEASUREMENTS OF DNA, RNA AND PROTEINS TO PERFORM COMPARATIVE ANALYSES OF MICROBIAL COMMUNITIES FROM THE ENVIRONMENTS

Abstract

ABSTRACT

Simple and rapid methods for the quantification of DNA, RNA and proteins using specific fluorescent dyes are proposed for the comparison and monitoring of microbial communities from the environment. The purpose of this study was the use of straightforward in situ methods which voided the need for preservation of samples and the risk of potential degradation and quantitative changes during transportation. Aside from this, methods used to obtain information on environmental microbial communities are generally time-consuming and present certain difficulty above all when working on solid substrates such as soils and rocks. New generation fluorescent dyes that bind specifically to DNA, RNA and proteins allow simple and rapid estimates of these biomolecules in crude environmental samples.

PRACTICAL APPLICATIONS

Monitoring the metabolic state of microbial communities on different substrates and environments is a requirement for comparing samples and assessing the participation of microorganisms in a variety of processes. Solid substrates are not easily analyzed by microscopic techniques and they require long processing times and tedious work. Aside from this, only a minor fraction (<1%) of microorganisms in most natural environments can be cultured in standard microbiological media (Ward et al. 1990). Other studies using incorporation of labeled substrates to approach activity rates or biomolecule extractions represent complex and long procedures during environmental studies.In order to evaluate microbial communities in a variety of substrates and environments, rapid and simple methods are proposed by measuring DNA, RNA and/or proteins using specific fluorescent dyes, without a need for prior purification, from crude solid samples.