Author/Authors :
khosravi babadi, zahra shahid beheshti university - faculty of life sciences and biotechnology - department of microbiology and microbial biotechnology, tehran, iran , khosravi babadi, zahra helmholtz centre for infection research gmbh (hzi) - microbial strain collection, braunschweig, germany , narmani, abolfazl university of tabriz - faculty of agriculture - department of plant protection, tabriz, iran , narmani, abolfazl helmholtz centre for infection research and german centre for infection research (dzif) - department of microbial drugs, partner site hannover/braunschweig, braunschweig, germany , ebrahimipour, gholam hossein shahid beheshti university - faculty of life sciences and biotechnology - department of microbiology and microbial biotechnology, tehran, iran , wink, joachim helmholtz centre for infection research gmbh (hzi) - microbial strain collection, braunschweig, germany , wink, joachim german centre for infection research association (dzif) - partner site hannover-braunschweig, braunschweig, germany
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.
Keywords :
actinobacteria , antimicrobial analyses , dna amplification technique , dna extraction method , dna–dna hybridization , whole genome sequencing