Generating the Starting Library

Random pools can also be generated by using various forms of mutagenic PCR (Section 8.3.1.4). The pools derived from these methods do not have the same 

The total yield should be 50pmol per 100 pL reaction after silica purification (Section 8.3.1.5). 

An alternative procedure for high-fidelity amplification is achieved by lowering the cINTP and Mg2+ concentrations. In addition, a higher-fidelity polymerase such as Pfu (PfuUltra, Stratagene) can be used. Yields are generally reduced, but this procedure has specific applications in cloning or characterization of catalysts with Ten own sequence, where mutations would impede analysis. 

Pfu DNA polymerase can be substituted for Taq DNA polymerase, but the manufacturer (Stratagene) recommends using the supplied buffer. 

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