Comparison of Five Methods for Extraction of Genomic Dna From a Marine Archaea, Pyrococcus Furiosus Publisher



Mirmohammadsadeghi H¹ ; Abedi D¹ ; Mohmoudpour HR¹ ; Akbari V¹ Show Affiliations
Source: Pakistan Journal of Medical Sciences Published:2013

Abstract

Objective: Archaea are one of the interesting forms of life due to their unique structural, physiological, biochemical and genetic features. For molecular analysis, high quality DNA should be extracted from these species. Methodology: In the present study, five extraction methods were evaluated for the isolation of DNA from Pyrococcus furiosus and the results compared. The DNA extracts were analyzed using gel electrophoresis and UV-spectophotometry. Results: Our results showed that a combination of enzymatic lysis (proteinase K) and chemical lysis (SDS) is the most suitable procedure for cell disruption of P. furiosus. This study showed that using freeze/thaw alone is ineffective in cell disruption. Recovering nucleic acids from cell lysates using silica-based DNA binding and solvent extractions was not suitable for DNA recovery and purification. Conclusions: The method proposed by Ramakrishnan and his coworker for isolation of genomic DNA from elemental sulfur-reducing hyper-thermophilic Archaea was successful for recovering DNA with good quantity and quality.