用于分析土壤微生物结构变化规律的变性梯度凝胶电泳条件研究

Denaturing Gradient Gel Electrophoresis( DGGE) Conditions Use to Analyze Soil Microbial Structure Variation Pattern

研究确定土壤微生物基因组DNA提取方法、PCR扩增条件、DGGE电泳条件,为进一步研究分析土壤中微生物结构变化规律提供理论依据。土壤微生物基因组DNA提取采用直接法和间接法进行比较; PCR扩增条件调整扩增体系、DGGE电泳条件调整变性剂范围,并对其结果进行比较分析。通过对DGGE电泳相关条件的研究,结果显示,土壤中粗基因组DNA采用直接法提取,然后进行纯化; PCR扩增体系中加入BSA,DGGE电泳系统组成中变性剂浓度范围为35%～55%。确定了土壤微生物基因组DNA提取方法、PCR扩增条件、DGGE电泳条件,为后续的相关研究提供理论依据。

Determination of soil microbial genome DNA extraction methods, PCR amplification conditions, DGGE （denaturing gradient gel electrophoresis） conditions were studied in order to provide reliable theoretical basis for further study on the analysis on the variation pattern of microbial community structure in soil. Soil microbial genomic DNA extraction were compared using direct method and indirect method; PCR amplification condition adjustment amplification system, DGGE conditions to adjust the denaturant range, and the results were compared with analyses. The results showed that by studying the whole conditions associated with DGGE, it was concluded that the crude genomic DNA was extracted by direct method and then carried out purification. In PCR amplification system BSA was added, and the concentration of denaturants in the components of DGGE system was at 35%-55%. Therefore, the soil microbial genome DNA extraction method, PCR amplification conditions, DGGE conditions were confirmed and provided real, reliable, accurate theoretical foundation for the follow up studies.