Chromosome sorting from hybrid cells offers enormous advantages for gene mapping and cloning, but purification of most chromosomes has been largely hindered by their similarity in size to other chromosomes. We have developed a novel cell line and strategy that allows simple, mass purification of mammalian chromosomes, permitting significant target genome enrichment. This strategy takes advantage of the small number of giant chromosomes (1,2,X) of the female Indian muntjac, a barking deer, avoiding the problem of size similarity. We introduced human chromosomes into a cell line derived from a muntjac and purified them to homogeneity using a relatively simple technique. This strategy should facilitate the isolation of chromosomes from species other than human for which hybrid cells are not available currently.