We have devised a general strategy for gene mapping based upon the direct amplification of a target sequence within a single microdissected Giemsa-banded chromosomal segment using the polymerase chain reaction. The usefulness of this approach was demonstrated by mapping a cloned human brain sodium channel (a subunit) gene sequence to chromosome 2q22-q23. When DNA from single, disected chromosome segments 2q21-qter and 2q24-pter were used as templates, a sodium channel-spedfic 172-base-pair polymerase chain reaction product was obtained. This product was not synthesized when segments 2q21-pter and 2q24-qter were used. Chromosome mic ionolymerase chain reaction is not only a simple, fast, and accurate method for gene mapping but also may offer sgiant advantages for other applications, such as cancer cytogenetics and linkage analysis.Assignment ofhuman genes or polymorphic DNA markers to specific chromosomal locations is a prerequisite for linkage analysis, location of individual genes, and the definition of their structure and function. In this communication we present a strategy for chromosome mapping that is direct, fast, and simple compared with traditional mapping techniques.Several methods, including somatic-cell genetics, in situ hybridization, and fluorescence-activated cell sorting of metaphase chromosomes, have been widely used to map structural genes and other identifiable DNA sequences onto chromosomes (1, 2). The most commonly used method for identifying the chromosomal locus of a specific human DNA sequence is in situ hybridization with radiolabeled probes. This approach is lengthy, labor-intensive, and often difficult to interpret because of cross-hybridization of the probe to related DNA sequences. Further, its mapping precision is limited by a variety of factors, including distortions in banding patterns caused by the preparation of specimens for hybridization and the spatial dispersion of isotopic signals captured on an emulsion overlay. Many of these drawbacks have been obviated by