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Synthetic DNA pp 129-138 | Cite as

The Polymerase Step Reaction (PSR) Method for Gene and Library Synthesis

  • Brian S. DeDeckerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1472)

Abstract

Current gene synthesis methods often incorporate a PCR-amplifying step in order to yield sufficient final product that is detectable and resolvable from multiple off-products. This amplification step can cause stochastic sampling effects that propagate errors during the synthesis and lower the variability when applied towards the construction of randomized libraries. We present the method for polymerase step reaction (PSR), a simple DNA polymerase-based gene synthesis reaction that assembles DNA oligonucleotides in a unidirectional fashion without the need for a PCR-type amplification (Lee et al., BioTechniques 59:163–166, 2015). The PSR method is simple and efficient with little off-product production, undetected stochastic sampling effects, and maximized variability when used to synthesize phage display libraries.

Key words

Gene synthesis Gene assembly Randomized library construction DNA polymerase Stochastic sampling effect Thermal cycling reaction Variable library and single-stranded DNA 

Notes

Acknowledgment

This work was supported in part by a grant from the National Institute of Health.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Molecular, Cellular and Developmental BiologyUniversity of ColoradoBoulderUSA

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