藏成药石榴健胃散基因组DNA提取方法的比较及优化

Comparison and optimization extraction of genomic DNA from Tibetan patent medicine Shiliu Jianwei powder

目的:比较不同试剂盒,不同方法对藏成药石榴健胃散基因组DNA的提取效果。方法:针对磁珠法与吸附柱法2类方法、4种不同市售试剂盒,延长裂解时间后,按照试剂盒标准操作方法,对藏成药石榴健胃散的基因组DNA进行提取,并通过Nanodrop one、Qubit、凝胶电泳检测综合比较各种方法对其基因组DNA提取的效果。结果:通过4种市售试剂盒、2类方法对藏成药基因组DNA的提取,结果显示吸附柱法即试剂盒3和试剂盒4提取的基因组DNA浓度、DNA片段完整性、PCR扩增效果方面优于磁珠法即试剂盒1和试剂盒2,且经济性更好。结论:初步探究了主流的核酸提取方法对藏成药石榴健胃散基因组DNA的提取效果,证明了试剂盒标准方法下,吸附柱法具有较好的基因组DNA提取效果,为后续进一步的提取方法的优化奠定了前期基础。

Objective:To compare the effects of different kits and different methods on the extraction of genomic DNA from Tibetan patent medicine Shiliu Jianwei powder.Methods:The genomic DNA of the Tibetan patent medicine Shiliu Jianwei powder was extracted using four different commercially available kits and magnetic bead method and the adsorption column method.The effect of genomic DNA extraction was comprehensively compared with Nano Drop one,Qubit,and gel electrophoresis.Results:Four commercially available kits and two methods were used to extract the genomic DNA of the Shiliu Jianwei powder.The results showed that the adsorption column method,namely kit 3 and kit 4,was superior than the bead method using kit 1 and kit 2 in terms of the concentration of genomic DNA,the integrity of DNA fragments,and the PCR amplification effect,and economy.Conclusion:Preliminary research on the extraction effect of main stream nucleic acid extraction methods ongenomic DNA of Tibetan patent medicine Shiliu Jianwei powder is performed.It is proved that under the standard method of the kit,the adsorption column method has a better extraction effect of genomic DNA,which lays a preliminary foundation for the further optimization of the extraction method.