The Establishment of the Real-Time PCR System for Detecting Soil Bacteria Abundance

For establishing the system of quantitative analysis for soil bacteria population,3 soil treatments including the sample P1 and P2 which was polluted by polycyclic aromatic hydrocarbon(PAH),and sample C from bare land were sampled and analyzed.Using the molecular cloning method,the standard substance of real-time PCR was structured,and the relevant parameter was optimized including the linearity(R²),amplification efficiency(E) and slope(S)of the standard curve.The results indicated that the relevant parameter is regular for real-time PCR,which R² was 0.985,E was 101.7% and S was 3.282.The quantity of bacteria cell in soil sample P1,P2 and C was quantitated as 1.4×10⁹,4.0×10⁹and 5.0×10⁹ respectively.It indicated that the abundance of soil bacteria could be quantitated by the established system of real-time PCR using unknown sensitive template.