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Simple and efficient modification of Golden Gate design standards and parts using oligo stitching

Jonas De Saeger (UGent) , Mattias Vermeersch (UGent) , Christophe Gaillochet (UGent) and Thomas B. Jacobs (UGent)
(2022) ACS SYNTHETIC BIOLOGY. 11(6). p.2214-2220
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Abstract
The assembly of DNA parts is a critical aspect of contemporary biological research. Gibson assembly and Golden Gate cloning are two popular options. Here, we explore the use of single stranded DNA oligos with Gibson assembly to augment Golden Gate cloning workflows in a process called "oligo stitching". Our results show that oligo stitching can efficiently convert Golden Gate parts between different assembly standards and directly assemble incompatible Golden Gate parts without PCR amplification. Building on previous reports, we show that it can also be used to assemble de novo sequences. As a final application, we show that restriction enzyme recognition sites can be removed from plasmids and utilize the same concept to perform saturation mutagenesis. Given oligo stitching's versatility and high efficiency, we expect that it will be a useful addition to the molecular biologist's toolbox.
Keywords
Gibson assembly, Golden Gate cloning, oligo stitching, pegRNA, saturation mutagenesis, synthetic biology, SYNTHETIC BIOLOGY

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MLA
De Saeger, Jonas, et al. “Simple and Efficient Modification of Golden Gate Design Standards and Parts Using Oligo Stitching.” ACS SYNTHETIC BIOLOGY, vol. 11, no. 6, 2022, pp. 2214–20, doi:10.1021/acssynbio.2c00072.
APA
De Saeger, J., Vermeersch, M., Gaillochet, C., & Jacobs, T. B. (2022). Simple and efficient modification of Golden Gate design standards and parts using oligo stitching. ACS SYNTHETIC BIOLOGY, 11(6), 2214–2220. https://doi.org/10.1021/acssynbio.2c00072
Chicago author-date
De Saeger, Jonas, Mattias Vermeersch, Christophe Gaillochet, and Thomas B. Jacobs. 2022. “Simple and Efficient Modification of Golden Gate Design Standards and Parts Using Oligo Stitching.” ACS SYNTHETIC BIOLOGY 11 (6): 2214–20. https://doi.org/10.1021/acssynbio.2c00072.
Chicago author-date (all authors)
De Saeger, Jonas, Mattias Vermeersch, Christophe Gaillochet, and Thomas B. Jacobs. 2022. “Simple and Efficient Modification of Golden Gate Design Standards and Parts Using Oligo Stitching.” ACS SYNTHETIC BIOLOGY 11 (6): 2214–2220. doi:10.1021/acssynbio.2c00072.
Vancouver
1.
De Saeger J, Vermeersch M, Gaillochet C, Jacobs TB. Simple and efficient modification of Golden Gate design standards and parts using oligo stitching. ACS SYNTHETIC BIOLOGY. 2022;11(6):2214–20.
IEEE
[1]
J. De Saeger, M. Vermeersch, C. Gaillochet, and T. B. Jacobs, “Simple and efficient modification of Golden Gate design standards and parts using oligo stitching,” ACS SYNTHETIC BIOLOGY, vol. 11, no. 6, pp. 2214–2220, 2022.
@article{8758708,
  abstract     = {{The assembly of DNA parts is a critical aspect of contemporary biological research. Gibson assembly and Golden Gate cloning are two popular options. Here, we explore the use of single stranded DNA oligos with Gibson assembly to augment Golden Gate cloning workflows in a process called "oligo stitching". Our results show that oligo stitching can efficiently convert Golden Gate parts between different assembly standards and directly assemble incompatible Golden Gate parts without PCR amplification. Building on previous reports, we show that it can also be used to assemble de novo sequences. As a final application, we show that restriction enzyme recognition sites can be removed from plasmids and utilize the same concept to perform saturation mutagenesis. Given oligo stitching's versatility and high efficiency, we expect that it will be a useful addition to the molecular biologist's toolbox.}},
  author       = {{De Saeger, Jonas and Vermeersch, Mattias and Gaillochet, Christophe and Jacobs, Thomas B.}},
  issn         = {{2161-5063}},
  journal      = {{ACS SYNTHETIC BIOLOGY}},
  keywords     = {{Gibson assembly,Golden Gate cloning,oligo stitching,pegRNA,saturation mutagenesis,synthetic biology,SYNTHETIC BIOLOGY}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{2214--2220}},
  title        = {{Simple and efficient modification of Golden Gate design standards and parts using oligo stitching}},
  url          = {{http://doi.org/10.1021/acssynbio.2c00072}},
  volume       = {{11}},
  year         = {{2022}},
}
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