The present study describes modifications to an existing bioautographic method for detecting genotoxic substances on thin-layer chromatography (TLC) plates by using the umu assay. It will be shown that the complex procedure that is described in the literature can be significantly simplified. Using the modified method, it is possible to detect 0.3 ng of the genotoxic substance 4-nitroquinoline-N-oxide. With 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal), a suitable substrate for environmental samples for the detection of the resulting β-galactosidase was found. Experiments showed, in contrast, that indirectly genotoxic substances could not thus far be identified on the TLC plate with overlayer bioautography.