In contrast to the library types described above, a variety of artificial methods exist for making libraries of variant genes. Variation throughout the gene can be introduced randomly by either error-prone PCR, DNA shuffling to recombine parts of similar genes together, or transposon-based methods to introduce indels.
Alternatively, mutations can be targeted to specific codons during de novo synthesis or saturation mutagenesis to construct one or more point mutants of a gene in a controlled way. This results in a mixture of double stranded DNA molecules which represent variants of the original gene.
The expressed proteins from these libraries can then be screened for variants which exhibit favorable properties (e.g. stability, binding affinity or enzyme activity). This can be repeated in cycles of creating gene variants and screening the expression products in a directed evolution process.