Fluorescence in situ hybridization (FISH) techniques were applied to the mapping of 18 probes from chromosome 18, permitting seven new assignments: D18S16 (18p11.31), D18S12 (18p11.1), D18S1 (18q12.2), D18S13 (18q21.31), D18S18 (18q21.31), D18S14 (18q21.33), and D18S17 (18q23). In addition, the localization of D18S3 in 18p11.3 was confirmed and that of D18S6, previously mapped in 18p11, was changed to 18q21.13. Finally, a more accurate mapping for nine probes was proposed: D18S7 (18q12.2), D18S10 (18q12.2), D18S24 (18q21.13), D18S8 (18q21.13), GRP(18q21.31), BCL2(18q21.33), D18S5(18q22.1), D18S19 (18q22.1), and D18S11 (18q22.3). The ordering of probes located in close proximity was made possible by combined use of single-color FISH with direct assignment on banded chromosomes, chromosomal length measurements, and double-color FISH.