Publisher Summary This chapter focuses on microcloning of mammalian metaphase chromosomes. Microcloning may be used to obtain libraries of clones from individually dissected mammalian chromosomes as well as chromosome regions. The numbers of chromosomes needed for linkage studies and for starts of chromosome walks are moderate, about 100–500 genome equivaleuts. Microcloning is also the method of choice when the amount of material is limited or where sufficient amounts of bulk isolated specific chromosomes cannot easily be obtained. With the currently used equipment for microdissection relatively coarse dissections can be made. For work with higher resolution it will be necessary to develop the manipulator—for example, with piezoelectric control, and television monitor. Another line to pursue is to investigate to what extent banding techniques can be combined with microcloning work. Such developments are highly desirable, among others in view of increasing needs for polymorphic DNA markers in human linkage studies.