1. Recursive construction and error correction of a simple combinatorial library.
Recursive construction and error correction of a simple combinatorial library. The recursive construction of six p53 variants is illustrated top to bottom: a diagram describes the shared (A, C, E in gray) and unique (B, D, colored) components of the target p53 combinatorial DNA library. A library construction protocol is computed, where target library sequences are recursively divided into shared and unique components and then further divided into basic oligonucleotide sequences, which are then synthesized conventionally. Gray oligos are shared by all library variants, and colored segments are used by variants with the corresponding colors. Oligos are recursively combined in vitro as shown to form the six target p53 variants. These variants were cloned and sequenced, and errors were identified (marked in red on top of clones). An error‐free minimal cut (the non‐faded part of the graph below the minimal cut black line) of the library construction graph was computed from only four error‐prone clones (variants 1, 3, 5 and 6). Error‐free segments out of these clones (delimited) were used as inputs for another iteration of the recursive reconstruction protocol, this time producing error‐free clones of all six target library members. Expected execution times for each step using standard equipment are shown on the left.
Gregory Linshiz et al. Mol Syst Biol 2008;4:191
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