Gene and library synthesis without amplification: polymerase step reaction (PSR)

BioTechniques
Zhuo-Bin LeeBrian S DeDecker

Abstract

Current gene synthesis methods often incorporate a PCR amplification step in order to yield final material sufficient for resolution from multiple off-products. These amplification steps can cause stochastic sampling effects that propagate errors in gene synthesis or decrease variability when applied to the construction of randomized libraries. We have developed a simple DNA polymerase-based gene synthesis reaction, polymerase step reaction (PSR), that assembles DNA oligonucleotides in a unidirectional fashion without the need for amplification. We demonstrate that PSR is efficient, with little off-product production, no detectable error propagation, and maximized variability in the synthesis of a phage display library.

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Methods Mentioned

BETA
phage display
PCR
PCA
electrophoresis

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