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为了建立土壤环境样品中细菌群落的定量分析体系,选用了多环芳烃污染土壤处理P1和P2,以及无多环芳烃污染的裸地生态系统土壤C等3种处理,利用构建克隆子的方法制作标准样品,并对实时荧光定量PCR标准曲线的线性(R2)、扩增效率(E)以及斜率(S)等相关参数进行了优化。结果表明,标准曲线的线性R2=0.985、扩增效率E=101.7%以及斜率S=-3.282,均符合参数符合要求。土壤样品P1、P2和C的细菌16SrDNA片段拷贝数为1.4×109个·g-1、4.0×109个·g-1和5.0×109个·g-1。说明建立的实时荧光定量PCR检测体系可以灵敏地定量未知模板的土壤细菌数量。图3,参11。
In order to establish a quantitative analysis system of bacterial community in soil environment samples, we selected three treatments of PAHs contaminated soil treatment P1 and P2 and non-PAH contaminated bare soil ecosystem C so as to make use of cloning method (R2), amplification efficiency (E), slope (S) and other related parameters of the real-time fluorescence quantitative PCR standard curve were optimized. The results showed that the linearity of the standard curve was R2 = 0.985, the efficiency of amplification was E = 101.7% and the slope was S = -3.282, both of which met the requirements of the parameters. The bacterial 16S rDNA fragment copy numbers of soil samples P1, P2 and C were 1.4 × 10 9 g -1, 4.0 × 109 g · g -1, and 5.0 × 109 g · g -1. The established real-time fluorescence quantitative PCR detection system can sensitively quantify the amount of soil bacteria in the unknown template. Figure 3, Senate 11.