We consider nonadaptive pooling designs for unique-sequence screening of a 1530-clone map of Aspergillus nidulans. The map has the properties that the clones are, with possibly a few exceptions, ordered and no more than 2 of them cover any point on the genome. We propose two subdesigns of the Steiner system S(3, 5, 65), one with 65 pools and approximately 118 clones per pool, the other with 54 pools and about 142 clones per pool. Each design allows 1 or 2 positive clones to be detected, even in the presence of substantial experimental error rates. Move efficient designs are possible if the overlap information in the map is exploited, if there is no constraint on the number of clones in a pool, and if no error tolerance is required. An information theory lower bound requires at least 12 pools to satisfy these minimal criteria, and an ''interleaved binary'' design can be constructed on 20 pools, with about 380 clones per pool. However, the designs with more pools have important properties of robustness to various possible errors and general applicability to a wider class of pooling experiments. (C) 1997 Academic Press.