Ligation-free gene synthesis by PCR: synthesis and mutagenesis at multiple loci of a chimeric gene encoding OmpA signal peptide and hirudin.

Abstract

A unique kination and ligation-free method that allows de novo synthesis of a gene through a novel application of polymerase chain reaction (PCR) involving stepwise elongation of sequence (SES) is described. SES-PCR is simple and efficient. Optimal utilization of nucleotides, ability to use only partially purified oligodeoxyribonucleotides, and elimination… (More)

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