The detection of a single photon from a laser interacting with an atomic ensemble is shown to produce entanglement of almost 3,000 atoms; in contrast to previous production of multi-atom entanglement, the highly non-classical nature of the present entangled state is verified by measurement of a negative quasiprobability distribution. Entanglement is the crucial resource that quantum technologies harness for classically impossible applications. Large ensembles of entangled particles promise to be useful in quantum metrology, but they are difficult to produce. Vladan Vuletic and colleagues demonstrate how the detection of a single photon from a laser interacting with an atomic ensemble can produce entanglement of almost 3,000 atoms. Entanglements of atomic ensembles with many atoms have been produced before, but they showed only weak signs of non-classicality, as quantified by the Wigner function. The entangled atoms described here, however, show a negative Wigner function and are thus highly non-classical, providing a realistic route to 'Schrodinger's cat' states in large atomic ensembles. The result has yet to be optimized to yield a quantum advantage in metrology applications but it is an important step towards this goal. Quantum-mechanically correlated (entangled) states of many particles are of interest in quantum information, quantum computing and quantum metrology. Metrologically useful entangled states of large atomic ensembles have been experimentally realized1,2,3,4,5,6,7,8,9,10, but these states display Gaussian spin distribution functions with a non-negative Wigner quasiprobability distribution function. Non-Gaussian entangled states have been produced in small ensembles of ions11,12, and very recently in large atomic ensembles13,14,15. Here we generate entanglement in a large atomic ensemble via an interaction with a very weak laser pulse; remarkably, the detection of a single photon prepares several thousand atoms in an entangled state. We reconstruct a negative-valued Wigner function—an important hallmark of non-classicality—and verify an entanglement depth (the minimum number of mutually entangled atoms) of 2,910 ± 190 out of 3,100 atoms. Attaining such a negative Wigner function and the mutual entanglement of virtually all atoms is unprecedented for an ensemble containing more than a few particles. Although the achieved purity of the state is slightly below the threshold for entanglement-induced metrological gain, further technical improvement should allow the generation of states that surpass this threshold, and of more complex Schrodinger cat states for quantum metrology and information processing. More generally, our results demonstrate the power of heralded methods for entanglement generation, and illustrate how the information contained in a single photon can drastically alter the quantum state of a large system.