RESUMEN

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.

Asunto(s)

ADN Polimerasa Dirigida por ADN/metabolismo , Biblioteca de Genes , ADN/genética , ADN/aislamiento & purificación , Cartilla de ADN , ADN de Cadena Simple/genética , ADN de Cadena Simple/aislamiento & purificación , ADN Polimerasa Dirigida por ADN/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa , Anticuerpos de Dominio Único/genética