Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis

• Published in: Baghdad journal of biochemistry & applied biological sciences 2022-06, Vol.3 (2), p.133-144
• Main Authors: Brito, Oslaene Alves de, Santos, Francisca Alves dos, Al Yafawi, Tassia Thais, Saraiva, Cícero Roberto Nascimento, Macedo da Silva, Rakel Olinda, Leandro, Lívia Maria Garcia, De Aquino, Pedro Everson Alexandre, De Sousa Júnior, Dárcio Luiz, Silva Leandro, Maria Karollyna do Nascimento
• Format: Article
• Language: English
• ISSN: 2706-9907; 2706-9915
• DOI: 10.47419/bjbabs.v3i02.123

Background and objective: Given the difficulty in establishing an ideal method that can successfully extract bacterial deoxyribonucleic acid (DNA) in Gram-negative bacteria, the objective of this work was to compare protocols for the extraction of bacterial DNA and to suggest more practical, faster, and less costly methodologies. Methods: Bacterial species used were Escherichia coli, provided by Dr. Leão Sampaio University Center. The evaluated/adapted methodologies were: sodium dodecyl sulfate (SDS); salting-out; Promega kit and cetyltrimethylammonium bromide (CTAB), and phenol/chloroform. Extracted DNA was examined by the agarose gel electrophoresis. Results: The SDS method revealed the best result in extracting DNA from E. coli. Its advantages include rapidness, low cost, and good concentration of the extracted material. Others methods showed low-quality DNA, probably due to the presence of large amounts of proteins in the cell wall of E. coli, interfering with the quality of the samples. Conclusions: It was concluded that the SDS method is better than others with better DNA quality, low cost, and good efficiency.