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Randomize Gene Sequences with New PCR Mutagenesis Kit

/TTP/dGTP), which leads to significant bias in Taqs mutational spectrum (Ts/Tv ratio, 2.7; ATGC/GCAT ratio, 10).1,2

GeneMorph Kit Provides Superior Yields

The GeneMorph kit must employ a robust PCR enzyme to ensure high product yield from low DNA template concentrations (highly mutagenic conditions). Since Mutazyme DNA polymerase is inherently error prone, high mutation frequencies are achieved using optimal PCR reaction conditions. In contrast, employing Mn2+ and unbalanced dNTP pools to lower the fidelity of Taq leads to reduced product yield,8 and such conditions are generally only useful for amplifying targets up to 1 kb in length.1

The superior performance of Mutazyme DNA polymerase is demonstrated in Figure 6. A series of targets, 0.65 kb to 10 kb in length, were amplified under error-prone conditions using Mutazyme and Taq DNA polymerases. For Taq reactions, the error-prone reaction conditions were employed,2 except that the MgCl2 concentration was decreased from 7 mM to 1.5 mM. PCRs were performed using identical cycling conditions and DNA template concentrations. Mutazyme DNA polymerase (2.5 U) produced high yields of all amplicons, while Taq DNA polymerase (5 U) successfully amplified only the shortest 650-bp product (Figure 6). The 650-bp product was not successfully amplified using 2.5 U Taq DNA polymerase or buffers containing 7 mM MgCl2, as recommended2 (data not shown).

Fig.6

Conclusions

Stratagenes GeneMorph PCR Mutagenesis kit is
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