JCVI: Oligonucleotide Assembly In Yeast to Produce Synthetic DNA Fragments.
Section Banner



Gibson DG

Oligonucleotide Assembly In Yeast to Produce Synthetic DNA Fragments.

Methods in molecular biology (Clifton, N.J.). 2012 Jun 01; 852: 11-21.

External Citation


The yeast Saccharomyces cerevisiae can take up and assemble at least 38 overlapping single-stranded oligonucleotides and a linear double-stranded vector in one transformation event. These oligonucleotides can overlap by as few as 20 bp and can be as long as 200 nucleotides in length to produce kilobase-sized synthetic DNA molecules. A protocol for designing the oligonucleotides to be assembled, transforming them into yeast, and confirming their assembly is described here. This straightforward scheme for assembling chemically synthesized oligonucleotides can be a useful tool for building synthetic DNA molecules.