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September 10, 2009Nucleic Acids Research297 citationsOpen Access

Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides

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DGDaniel G. Gibson

Key Points

  • This research aims to showcase a method for assembling DNA fragments using overlapping oligonucleotides in yeast.
  • Utilized the yeast Saccharomyces cerevisiae for DNA assembly
  • Involved transforming 38 overlapping single-stranded oligonucleotides with a linear double-stranded vector in one event
  • Oligonucleotides varied from 20 bp overlaps to lengths of 200 nucleotides
  • Successfully assembled DNA fragments in one transformation event
  • Demonstrated efficient uptake of multiple overlapping oligonucleotides
  • Proposed a straightforward scheme for constructing synthetic DNA molecules

Abstract

Here it is demonstrated that 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. This straightforward scheme for assembling chemically-synthesized oligonucleotides could be a useful tool for building synthetic DNA molecules.

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Cite This Study

Daniel G. Gibson (2009) studied this question.

synapsesocial.com/papers/6a10c5bdacd1dbe064646176https://doi.org/10.1093/nar/gkp687
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