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sub> concentration. Under these conditions, Taq preferentially introduces AT, TA, AG, and TC mutations (66%-68% of total mutations) and rarely incorporates GC and CG transversions. In comparison, Mutazyme DNA polymerase tends to produce GA, CT, GT, and CA changes (64% total mutations), and all transversion mutations are represented. Taq is approximately three times more likely to mutate As and Ts than Gs and Cs (ATGC/GCAT ratio >1^2,3), while Mutazyme DNA polymerase is nearly three times more likely to mutate Gs and Cs (ATGC/GCAT ratio 0.2). Both enzymes prefer to incorporate transition mutations rather than transversion mutations (Ts/Tv = 0.8-1.2) (Table 2).

As shown in Table 2, Mutazyme DNA polymerase produces a broad and unique spectrum of mutations, thereby facilitating the production of representative mutant libraries. The distinct mutational spectra of Mutazyme and Taq DNA polymerases implies that different types of mutations will be present in mutant libraries constructed by these different error-prone methods. For example, the GeneMorph kit facilitates mutagenesis of GC-rich targets and synthesis of GCCG transversions, both of which are relatively difficult for Taq DNA polymerase. Since mutation levels are varied without altering reaction conditions, the broad spectrum of mutations produced by the GeneMorph kit is maintained over a wide range of mutation frequencies, even at high mutation levels (6-13 mutations per kb/2 PCRs). In contrast, to achieve high mutation frequencies with Taq (>4.9-6.6 mutations per kb^2,3), it is typical to increase the dGTP concentration (final conditions: 0.5 mM MnCl₂; 0.2 mM dATP; 1 mM dCTP
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