Heat Treatment of Bacteria: A Simple Method of DNA Extraction for Molecular Techniques

Abstract

Objective: To evaluate the efficacy of two simple methods involving use of heat for extraction of bacterial deoxyribonucleic acid (DNA) be used in molecular techniques like polymerase chain reaction (PCR), restriction fragments length polymorphism (RFLP) and DNA sequencing and compare them with DNA extraction using commercial kits. Design: DNA extraction by improved alternative methods and commercial kit. Setting: Microbiology Research Laboratory, Faculty of Allied Health Sciences, Kuwait University, Kuwait Material: Forty isolates of Klebsiella pneumoniae Intervention: DNA was extracted from isolates by either boiling for 10 minutes or microwave irradiation for 10 seconds. For comparison, DNA was also extracted using a commercial kit. All extracted DNA samples were analyzed by PCR, RFLP and / or DNA sequencing of TEM and SHV genes of the bacteria. Main Outcome Measures: Successful extraction of DNA Results: PCR, RFLP and DNA sequencing gave the expected results in all the DNA samples extracted by all the three methods (boiling, microwave irradiation and the commercial kit). The results were qualitatively equivalent in all methods. Conclusion: Heat may be used to extract DNA from K. pneumoniae which can be utilized successfully in performing PCR, RFL and DNA sequencing.