In order to efficiently select aptamers that bind to and inhibit proteins, we developed a method that involves screening DNA aptamers based on their inhibitory activities using an evolution-mimicking algorithm after the pre-selection by SELEX. The value of this method was demonstrated by the identification of an inhibitor of Taq DNA polymerase in a unique single-stranded DNA library, which was expected to form a G-quartet structure. This method consists of selection via an inhibition assay, sequence shuffling, and mutation in silico. After six rounds of selection, the inhibitory activities of the aptamers had evolved significantly. This demonstrates the utility of this strategy for screening aptamers based on their inhibitory actions. (c) 2006 Elsevier Inc. All rights reserved.