Cytogenetic methods for the mouse: preparation of chromosomes, karyotyping, and in situ hybridization

The ability to examine the chromosomes of murine cells is rapidly becoming essential to the mouse molecular geneticist. Although these technologies exist, the protocols are scattered throughout the literature, and in some cases different protocols exist for the same procedure. We attempt here both to augment and to consolidate this body of work. This review provides a complete set of procedures for manipulating mouse chromosomes. Prospective sources of metaphase chromosomes are discussed and detailed protocols are outlined for their preparation and characterization. We assume that the reader has little or no cytogenetic background and have included a troubleshooting guide to assist the process. Protocols for in situ hybridization to metaphase spreads that allow the rapid localization of sequences using nonradioactive probes are also included. Variations in the protocols described here abound and alternatives are indicated.