use of real-time PCR technology to assess the effectiveness of methods of DNA extraction from cultures of acidophilic chemolithotrophic microorganisms

• Published in: Applied biochemistry and microbiology Vol. 47; no. 2; pp. 206 - 210
• Main Authors: Rogatykh, S. V, Dokshukina, A. A, Khainasova, T. S, Muradov, S. V, Kofiadi, I. A
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : SP MAIK Nauka/Interperiodica 01-03-2011; SP MAIK Nauka/Interperiodica; Springer Nature B.V
• Subjects: Biochemistry; Biomedical and Life Sciences; Biotechnology; Chloroform; Deoxyribonucleic acid; DNA; Enzymes; Life Sciences; Medical Microbiology; Microbiology; Microorganisms; Phenols; Polymerase chain reaction; Real time; Kamchatka Peninsula; Lysis Activity; Apply Biochemistry; Archaea; Acidithiobacillus Thiooxidans
• ISSN: 0003-6838; 1608-3024
• DOI: 10.1134/S0003683811020177

Comparative evaluation of efficiency of several methods of DNA extraction from storage cultures of acidophilic chemolithotrophic microorganism communities isolated from sulfide ores of Shanuch ore deposit (Kamchatka peninsula) was conducted. DNA extraction methods in various combinations of physical (heating to 65-98°C, grinding with SiO₂ particles), enzymatic (treatment with lysozyme and proteinase K), and chemical (GuSCN, CTAB and KOH) treatments were tested. The evaluation of efficiency was performed using Real-time PCR. The best result was obtained for the combined method based on GuSCN lysis activity (lysis at 65°C) followed by purification with phenol and chloroform.