Techniques to facilitate controlled interactions between single photons and atoms are now being actively explored. These techniques are important for the practical realization of quantum networks, in which multiple memory nodes that utilize atoms for generation, storage and processing of quantum states are connected by single-photon transmission in optical fibres. One promising avenue for the realization of quantum networks involves the manipulation of quantum pulses of light in optically dense atomic ensembles using electromagnetically induced transparency (EIT, refs 8, 9). EIT is a coherent control technique that is widely used for controlling the propagation of classical, multi-photon light pulses in applications such as efficient nonlinear optics. Here we demonstrate the use of EIT for the controllable generation, transmission and storage of single photons with tunable frequency, timing and bandwidth. We study the interaction of single photons produced in a 'source' ensemble of 87Rb atoms at room temperature with another 'target' ensemble. This allows us to simultaneously probe the spectral and quantum statistical properties of narrow-bandwidth single-photon pulses, revealing that their quantum nature is preserved under EIT propagation and storage. We measure the time delay associated with the reduced group velocity of the single-photon pulses and report observations of their storage and retrieval.