The queen emerges from her cell after 15 days of development and she remains in the hive for 3–7 days before venturing out on a mating flight. Mating flight is otherwise known as "nuptial flight". Her first orientation flight may only last a few seconds, just enough to mark the position of the hive. Subsequent mating flights may last from 5 minutes to 30 minutes, and she may mate with a number of male drones on each flight. Over several matings, possibly a dozen or more, the queen receives and stores enough sperm from a succession of drones to fertilize hundreds of thousands of eggs. If she does not manage to leave the hive to mate—possibly due to bad weather or being trapped in part of the hive—she remains infertile and becomes a drone layer, incapable of producing female worker bees. Worker bees sometimes kill a non-performing queen and produce another. Without a properly performing queen, the hive is doomed.

Mating takes place at some distance from the hive and often several hundred feet in the air; it is thought that this separates the strongest drones from the weaker ones, ensuring that only the fastest and strongest drones get to pass on their genes.