CHEMICAL COMPOSITION OF ENVIRON FOR ISOLATION OF DNA FROM CELLS OF CHEMOLITHOTROPHIC MICROORGANISMS
The article describes the main commercial and experimental techniques for isolating DNA from microbial cells. The reliability of the molecular biological methods used in the analysis of the structure of microorganism communities depends on the quality and representativeness of the DNA preparation obtained in the process of isolation from the biomaterial. The optimal method of DNA isolation should provide effective washing of attached bacterial cells from soil particles, cell lysis and purification from inhibitors. Several combined DNA extraction protocols based on different types of effects on eubacterial and archaic cells have been compared experimentally. Methods based on physical (grinding of cells with SiO2 particles), enzymatic (treatment with proteinase K and lysozyme), chemical (treatment with GuSCN and CTAB), exposure to cells and a combined technique based on alkaline lysis (KOH) in the presence of a low molecular weight polyethylene glycol polymer PEG-200). It is recommended to use a technique based on the lysing activity of guanidinium isothiocyanate for molecular analysis of the structure of the communities of acidophilic chemolithotrophic microorganisms.