革兰氏阳性细菌基因组DNA提取方法的比较及优化

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

摘要:

【目的】基因组DNA提取效率和质量对分子生物学相关研究起着关键的作用，革兰氏阳性细菌由于细胞壁较厚、难破裂使其基因组DNA提取的难度增大，本文旨在寻找一种高效稳定的DNA提取方法。【方法】以Clostridium thermocellum和Thermoanaerobacterium thermosaccharolyticum为实验菌株，使用6种DNA提取方法对C. thermocellum基因组DNA进行提取，对比其提取效果和产率。【结果】改良的SDS-碱裂解法提取得到的DNA浓度较高(400 mg/l左右)，且平行样间浓度和纯度稳定。【结论】为革兰氏阳性细菌基因组DNA提取提供参考。

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.