Comparative Study of Extracted Nucleic Acid from Escherichia coli by Two Methods Phenol-chloroform and Extraction Using Magnetic Nanoparticle

摘要

Background: The most important researches in molecular and genetic engineering fields are the finding of optimal methods for extraction of the genomic content of microorganisms and cells using them, can achieve the most amount and purity with the least time and cost. There are several methods for the extraction of bacterial DNA and the use of magnetic nanoparticles is one of the novel methods of nucleic acid isolation. The aim of this study is to compare DNA extracted from Escherichia coli using two techniques including phenol-chloroform and magnetic nanoparticles. Methods: In this study, Escherichia coli strain ATCC25922 was used to compare the two extraction methods. For genomic DNA extraction the phenol-chloroform methods as well as the technique of using bilayer SiO2/Fe3O4 magnetic nanoparticles were used. Both products were analyzed by nanodrop and following agarose gel electrophoresis. Results: The results of optical density (OD) evaluation with nanodrop spectrophotometer showed that the concentration of extracted DNA with conventional phenol-chloroform method and magnetic nanoparticle extraction technique were 761.4 μg/ml and 531 μg/ml respectively. Conclusion: According to the obtained results by comparing the optical absorption of the two products and comparative evaluation with control groups in magnetic nanoparticle extraction method, also by comparing different factors such as the amount of time required, costs, ease of use, it can be concluded that due to the time-consuming and dangerous nature of the phenol-chloroform method, the using magnetic nanoparticles is an appropriated method for extracting the genomic content.