Comparison and optimization of methods for genomic DNA extraction from Gram positive bacteria

Abstract:

[Objective] The quality and efficiency of genomic DNA extraction play a key role in molecular biology research. The cell wall of Gram positive bacterium is thick and hard to be destroyed, which makes it difficult for genomic DNA extraction. The objective of this study was to find an efficient and repeatable method to extract the genomic DNA from Gram positive bacterium. [Methods] The genomic DNA of Clostridium thermocellum and Thermoanaerobacterium thermosaccharolyticum were extracted using six extraction methods. The DNA concentration and DNA purity were compared for these methods. [Results] The results showed that the concentration of DNA was highest (about 400 mg/L) and the consistency of DNA concentration and DNA purity were best when improved SDS alkaline lysis method was used. [Conclusion] This research provided a reference for the extraction of genomic DNA from Gram positive bacterium.