METHOD OF ISOLATION OF GENOME DNA FROM MICROORGANISM CELLS

摘要

molecular biology, microbiology. SUBSTANCE: invention relates to methods of isolation of DNA. Yeast or gram-positive microorganisms are treated with buffer solution of neutral pH value containing chaotrop as salt. Salt is taken among the group of guanidinium derivatives in concentration 4-8 M. Cells are incubated in extractant buffer at temperature by 5-10 C lower of DNA melting point in this buffer. Then genome DNA bound firmly with microorganism cellular envelopes is isolated followed by hydrolysis of components of microorganism cellular walls by lytic enzymes. Deproteinization and purification of DNA from proteins, lipids and polysaccharides and low-molecular components are carried out without preliminary disruption of cellular envelopes. Invention can be used in laboratory and research practice for isolation of high-molecular DNAs or their fragments (macrorestricts) from yeast cells and gram-positive microorganisms with high-strength cellular envelope that can be used then for analysis by pulse-electophoresis procedure, cloning and polymerase chain reaction carrying out. EFFECT: simplified procedure, decreased labor intensity, decreased cost of method. 2 cl