Abstract
We described a SYBR green-based quantitative PCR (qPCR) as an approach to estimate the relative abundances of Eubacteria and Archaea in methane fermentation process with soybean curd residue as substrates. Eubacterial and Archaeal primers were selected according the specificity and performance in qPCR firstly, then, optimization of three parameters (forward and reverse primer concentration, annealing temperature) was attempted using a Box–Behnken design under the response surface methodology (RMS) for enhancing the accuracy and sensitivity of qPCR. Quantitative PCR with the optimal conditions was applied for monitoring of microbial population dynamics during methane fermentation process.
