Microtubules are intrinsically polar structures. A consequence of this polarity is that the two ends of the microtubule polymer exhibit different properties. The more dynamic plus ends and the mechanisms that regulate their behavior have been the focus of much recent attention. Here, we concentrate on the dynamics and regulation of minus ends, which play distinct but equally critical roles in microtubule function. In the first part of this review, we compare the in vitro and in vivo behavior of microtubules from a minus end perspective. This comparison suggests that cells possess conserved mechanisms to specifically inhibit minus end polymerization, and perhaps also to actively promote depolymerization. In the second part, we focus on the spatial positioning of minus ends, which is achieved by localized microtubule nucleation, minus end capping and minus end anchoring as well as by motor-dependent sorting. These mechanisms are used in different biological contexts to generate the diversity of organized microtubule arrays in cells.