Extended Data Figure 1: DNAzyme cassette architectures with only one DNAzyme or with one class II and one class I enzyme are not suitable for staple production.

a, Schematic representation of a pseudogene in which adjacent staples are separated by a single DNAzyme, yielding staples with a 40-base-long DNAzyme overhang at their 3′ ends. b, Schematic representation of a pseudogene in which staples are separated by one class II and one class I DNAzyme, yielding staples without 3′ overhangs. c, d, Agarose-gel-electrophoretic characterization of the self-cleavage reaction of a pseudogene using the DNAzyme cassette architecture shown in b; c, self-cleavage at different salt concentrations at 37 °C; d, self-cleavage at different temperatures at salt concentrations of 5 mM MgCl₂ and 2 mM ZnSO₄. sta, reference sample containing chemically synthesized staples; DZ, reference sample containing chemically synthesized oligonucleotides with the sequence of the ‘waste’ DNAzymes.