Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 14 2 2022 04 13 186 193 Zahra Khosravi Babadi Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University GC, Tehran, Iran; Microbial Strain Collection, Helmholtz Centre for Infection Research GmbH (HZI), Braunschweig, Germany Abolfazl Narmani Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran; Department of Microbial Drugs, Helmholtz Centre for Infection Research and German Centre for Infection Research (DZIF), Partner Site Hannover/Braunschweig, Braunschweig, Germany Gholam Hossein Ebrahimipour Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University GC, Tehran, Iran Joachim Wink Microbial Strain Collection, Helmholtz Centre for Infection Research GmbH (HZI), Braunschweig, Germany; German Centre for Infection Research Association (DZIF), Partner site Hannover-Braunschweig, Braunschweig, Germany 2021 03 02 2022 03 11 Background and Objectives: DNA extraction is an important step of any molecular experiment. DNA could not be easily extracted from members of actinomycetes by the usual methods of lysis. Due to the low efficiency of the conventional DNA extraction methods, development of an effective technique for DNA extraction of actinobacteria in emergency cases seems to be necessary. Since most of the DNA extraction techniques and commercial kits are not efficient enough to extract DNA from actinobacteria, this study was conducted to improve an efficient method obtained from conventional one to extract DNA from this group of bacteria. Materials and Methods: DNA extraction was performed using five methods (an improved method, Invisorb Spin Plant Mini Kit, EZ-10 Spin Column, Sarrbrucken method (HZI, Germany) and Kirby Bauer's method). To evaluate the quantity and quality of extracted genomic DNA, UV absorbance of all samples and efficiency of polymerase chain reaction (PCR) were evaluated. Results: Overall, the results revealed that the highest quantity (up to 4000 ng/µl) and good quality of DNA was obtained using introduced DNA extraction method. Conclusion: Results indicated that recently introduced improved method was more efficient for extraction of DNA from actinobacteria for DDH (DNA–DNA hybridization) test and for those require the high concentration of DNA. https://ijm.tums.ac.ir/index.php/ijm/article/view/3019 https://ijm.tums.ac.ir/index.php/ijm/article/download/3019/1443