Leading scientists in cell biology, genetics, cell culturing, and gene transfer present their most productive methods for the preparation, characterization, and use of artificial chromosomes. The authors cover all relevant areas of artificial chromosome research—from basic genetics to daring attempts at building new gene therapy tools—and describe possible applications in genetic manipulation requiring large portions of the genome. Each fully tested protocol is described in step-by-step detail and includes a background introduction outlining the principle behind the technique, reagent and equipment lists, and tips on troubleshooting and how to avoid pitfalls.