Combination and improvement of conventional DNA extraction methods in Actinobacteria to obtain high-quantity and high-quality DNA

Zahra Khosravi Babadi; Abolfazl Narmani; Gholam Hossein Ebrahimipour; Joachim Wink

doi:10.18502/ijm.v14i2.9187

Zahra Khosravi Babadi Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University GC, Tehran, Iran
Abolfazl Narmani Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
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

DOI: https://doi.org/10.18502/ijm.v14i2.9187

Keywords: Actinobacteria; Antimicrobial analyses; DNA amplification technique; DNA extraction method; DNA–DNA hybridization; Whole genome sequencing

Abstract

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.