Bead-beating and isolation of environmental nucleic acids.
|Year of publication||2015|
|MU Faculty or unit|
|Description||Microorganisms represent fundamental and irreplaceable part of all ecosystems. They are responsible for nutrient cycling in nature and involved in nearly all important processes in terrestrial and aquatic ecosystems. They generate nutrients in bases of all food chains and their extensive metabolic potential is utilized in many areas of human activities. Further expansion of utilization of microbes or their products in agriculture, various biotechnological applications and other human activities depends mainly on increasing knowledge of their diversity in various ecosystems, mainly in soil, which represents immense source of microbial diversity. Various methods are used to approach and explore this diversity, majority of them being based on isolation of microbial nucleic acids. One of the first steps in all extraction methods is mechanical treatment of soil (and other) samples. Tone of the most widely used mechanical treatments these days is so-called bead-beating. This treatment serves physical disruption to disperse soil aggregates and – at the same time –starts the extraction process itself. It is performed using various instruments called bead-beater, ribolyser, MagNa Lyser etc, which all use vigorous shaking of samples in a buffer with various beads. It is surprising that not very big attention is payed to exact parameters of this treatment as it can utilize not only various types of beads, but can differ also in length and intensity of beating itself. We tested all these variables and their possible influence of the overall results of diversity analysis will be discussed in our presentation.|