Batch trials were carried out to evaluate the Biochemical Methane Potential (BMP) of 61 different substrates collected from agricultural farms and industrial sites in Estonia. Tests were performed in 500 mL plasma bottles at 36°C. The highest methane yield from all tested substrates was obtained from unconsumed dairy products (557 ± 101 L kg-1 VS) while the lowest was obtained from animal slurries (238 L kg-1 VS ± 42). From tested energy crops, foxtail millet achieved the highest methane yield (320 L kg-1 VS). Silages from different crops presented methane yields from 296 ± 31 L CH4 kg-1 VS to 319 ± 19 L CH4 kg-1 VS. The influence of chemical composition and kinetic rate constants (k) on methane potential was analyzed. Anaerobic digestibility of selected agro-industrial substrates was markedly influenced by their organic content, i.e. total proteins and lignin concentrations. Rate constants were found to correlate negatively with hemicellulose, cellulose and lignin (p < 0.05). Results from this study suggest that an appropriate characterization of the chemical composition of the substrates is important not only for predicting BMP and the kinetics rates, but also for identifying possible inhibitors during the anaerobic digestion process. Results on the BMP and national availability of studied substrates indicate that herbal biomass and agro-industrial residues are promising substrates for biogas production in agricultural biogas facilities in Estonia. © 2017, Eesti Pollumajandusulikool. All rights reserved.