The recovery of the grey wolf in Yellowstone National Park is an outstanding example of a successful reintroduction. A general question concerning reintroduction is the degree to which genetic variation has been preserved and the specific behavioural mechanisms that enhance the preservation of genetic diversity and reduce inbreeding. We have analysed 200 Yellowstone wolves, including all 31 founders, for variation in 26 microsatellite loci over the 10-year reintroduction period (1995–2004). The population maintained high levels of variation (1995 H0 = 0.69; 2004 H0 = 0.73) with low levels of inbreeding (1995 FIS = –0.063; 2004 FIS = –0.051) and throughout, the population expanded rapidly (N1995 = 21; N2004 = 169). Pedigree-based effective population size ratios did not vary appreciably over the duration of population expansion (1995 Ne/Ng = 0.29; 2000 Ne/Ng = 0.26; 2004 Ne/Ng = 0.33). We estimated kinship and found only two of 30 natural breeding pairs showed evidence of being related (average r = –0.026, SE = 0.03). We reconstructed the genealogy of 200 wolves based on genetic and field data and discovered that they avoid inbreeding through a wide variety of behavioural mechanisms including absolute avoidance of breeding with related pack members, male-biased dispersal to packs where they breed with nonrelatives, and female-biased subordinate breeding. We documented a greater diversity of such population assembly patterns in Yellowstone than previously observed in any other natural wolf population. Inbreeding avoidance is nearly absolute despite the high probability of within-pack inbreeding opportunities and extensive interpack kinship ties between adjacent packs. Simulations showed that the Yellowstone population has levels of genetic variation similar to that of a population managed for high variation and low inbreeding, and greater than that expected for random breeding within packs or across the entire breeding pool. Although short-term losses in variation seem minimal, future projections of the population at carrying capacity suggest significant inbreeding depression will occur without connectivity and migratory exchange with other populations.