In vitro reconstitution experiments have played an essential role in a large body of research on SNARE-mediated membrane fusion. We and others have developed single vesicle assays to gain more detailed insight into the kinetics of vesicle docking and fusion. Previously, we used supported membranes in combination with total internal reflection microscopy (TIRFM) to record the docking and fusion of vesicles containing recombinant Synaptobrevin2 (Syb2) to an acceptor SNARE complex consisting of one Syntaxin 1a (Syx1a), one SNAP25 (SN25) and a short Syb2 fragment (Syb49-96) to ensure a 1:1 stoichiometry between Syx1a and SN25.Here we report experiments in which the Syb2 containing vesicles were replaced by synaptic vesicles (SV) purified from rat brain. Docking and fusion of synaptic vesicles with supported membranes is efficient, fast (∼35 ms) and SNARE dependent. The fusion kinetics obtained with different lipid environments in the supported membrane and under different salt conditions are compared to the kinetics measured with reconstituted Syb2 vesicles.