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25,26 Furthermore, a polymorphism of the DHFR enzyme resulting in a 19bp deletion within the site of a key regulatory sequence has been theorized to exhibit negative effects on the expression and activity of this enzyme
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The three libraries were generated using error-prone PCR as described above, using the following conditions to produce varying levels of mutagenesis: Library 1: 30 M 8-oxo-dGTP, 1 M dPTP, 20 PCR cycles Library 2: 20 M 8-oxo-dGTP, 2 M dPTP, 20 PCR cycles Library 3: 10 M 8-oxo-dGTP, 3 M dPTP, 20 PCR cycles The library sizes (approximated by number of transformants as described above) were 6 10 7 for each library
