Evaluation Of three methods of DNA extraction from Pseudomonas aeruginosa by Spectrophotometer nanodrop and gel electrophoresis Evaluation Of three methods of DNA extraction from Pseudomonas aeruginosa by Spectrophotometer nanodrop and gel electrophoresis

Abstract

DNA extraction is the first stage of gene structure and function study, and finding an effective protocol for isolation DNA in genetic engineering and microbiology studies is essential. There are many different methods for DNA extraction which are interchangeable depending on the wall and membrane structure in the bacterium. Objective: The aim of this study was the assessment of different methods for DNA extraction from gram-negative bacterium (Pseudomonas aeruginosa) and the selection of effective protocol.Materials and methods: In this study three different methods of DNA extraction from gram-negative bacterium (Pseudomonas aeruginosa) is compared to determine the quantity and the quality of the DNA. These methods include lysis buffer containing proteinase K, DNGTM-plus kit and boiling. In each method, ten samples were selected in a completely random plan and the concentration of DNA was measured using spectrophotometer, Nanodrop and agarose gel electrophoresis. Result: Variance analysis illustrated a meaning difference between three methods. Also average comparison using Duncan test illustrated meaning difference. Finally, boiling method was considered as the most suitable method with the highest extraction efficiency.

Keywords

• DNA extraction
• spectrophotometer nanodrop
• gel electrophoresis
• variance analysis
• Duncan test