ArticleNucleic acids research2012
Error correction of microchip synthesized genes using Surveyor nuclease.
Article in Nucleic acids research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- POSoligo software for in vitro gene synthesis.Scientific reports · 2024Article
- Uncertainties in synthetic DNA-based data storage.Nucleic acids research · 2021Review
- Experiment and Simulation Reveal How Mutations in Functional Plasticity Regions Guide Plant Monoterpene Synthase Product Outcome.ACS catalysis · 2019Article
- Synthetic biology advances and applications in the biotechnology industry: a perspective.Journal of industrial microbiology & biotechnology · 2018Article
- DNA assembly with error correction on a droplet digital microfluidics platform.BMC biotechnology · 2018Article
- High-throughput construction of multiple cas9 gene variants via assembly of high-depth tiled and sequence-verified oligonucleotides.Nucleic acids research · 2018Article
- Highly selective retrieval of accurate DNA utilizing a pool of in situ-replicated DNA from multiple next-generation sequencing platforms.Nucleic acids research · 2018Article
- A systematic comparison of error correction enzymes by next-generation sequencing.Nucleic acids research · 2017Article
- High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment.Scientific reports · 2017Article
- High-throughput expression of animal venom toxins in Escherichia coli to generate a large library of oxidized disulphide-reticulated peptides for drug discovery.Microbial cell factories · 2017Article
- Synthetic DNA Synthesis and Assembly: Putting the Synthetic in Synthetic Biology.Cold Spring Harbor perspectives in biology · 2017Review
- Next generation gene synthesis: From microarrays to genomes.Engineering in life sciences · 2017Review
- Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently.Chemical Society reviews · 2015Review
- Synthetic Biology: A Bridge between Artificial and Natural Cells.Life (Basel, Switzerland) · 2014Review
- SpeedyGenes: an improved gene synthesis method for the efficient production of error-corrected, synthetic protein libraries for directed evolution.Protein engineering, design & selection : PEDS · 2014Article
- Error removal in microchip-synthesized DNA using immobilized MutS.Nucleic acids research · 2014Article
- Large-scale de novo DNA synthesis: technologies and applications.Nature methods · 2014Review
- 'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules.Nucleic acids research · 2012Article
- DNA synthesis, assembly and applications in synthetic biology.Current opinion in chemical biology · 2012Review
- Error correction in gene synthesis technology.Trends in biotechnology · 2012Review
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
The development of economical and high-throughput gene synthesis technology has been hampered by the high occurrence of errors in the synthesized products, which requires expensive labor and time to correct. Here, we describe an error correction reaction (ECR), which employs Surveyor, a mismatch-specific DNA endonuclease, to remove errors from synthetic genes. In ECR reactions, errors are revealed as mismatches by re-annealing of the synthetic gene products. Mismatches are recognized and excised by a combination of mismatch-specific endonuclease and 3'→5' exonuclease activities in the reaction mixture. Finally, overlap extension polymerase chain reaction (OE-PCR) re-assembles the resulting fragments into intact genes. The process can be iterated for increased fidelity. With two iterations, we were able to reduce errors in synthetic genes by >16-fold, yielding a final error rate of ∼1 in 8700 bp.
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Registered trials
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