ArticleNucleic acids research2005
Removal of mismatched bases from synthetic genes by enzymatic mismatch cleavage.
Article in Nucleic acids research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 63 citations in OpenAlex.
- Engineered eMutS based magnetic separation system for high-fidelity DNA synthesis.Synthetic and systems biotechnology · 2026Article
- Purification of multiplex oligonucleotide libraries by synthesis and selection.Nature biotechnology · 2022Article
- DNA assembly with error correction on a droplet digital microfluidics platform.BMC biotechnology · 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
- Multiplex pairwise assembly of array-derived DNA oligonucleotides.Nucleic acids research · 2016Article
- Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently.Chemical Society reviews · 2015Review
- 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
- Gene Assembly from Chip-Synthesized Oligonucleotides.Current protocols in chemical biology · 2012Article
- Error correction in gene synthesis technology.Trends in biotechnology · 2012Review
- Error correction of microchip synthesized genes using Surveyor nuclease.Nucleic acids research · 2012Article
- Discovery of T cell antigens by high-throughput screening of synthetic minigene libraries.PloS one · 2012Article
- High-fidelity gene synthesis by retrieval of sequence-verified DNA identified using high-throughput pyrosequencing.Nature biotechnology · 2010Article
- Genome engineering.Nature biotechnology · 2009Review
- Experimental analysis of gene assembly with TopDown one-step real-time gene synthesis.Nucleic acids research · 2009Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The success of long polynucleotide de novo synthesis is largely dependent on the quality and purity of the oligonucleotides used. Generally, the primary product of any synthesis reaction is directly cloned, and clones with correct products have to be identified. In this study, a novel strategy has been established for removing undesired sequence variants from primary gene synthesis products. Single base-pair mismatches, insertions and deletions were cleaved with specific endonucleases. Three different enzymes--T7 endonuclease I, T4 endonuclease VII and Escherichia coli endonuclease V--have been tested. As a model, a synthetic polynucleotide encoding the bacterial chloramphenicol-acetyltransferase (cat) was synthesized using different methods for one step polynucleotide synthesis based on ligation of oligonucleotides. The influence of enzymatic mismatch cleavage (EMC) as an error correction step on the frequency of correct products was analyzed by functional cloning of the synthetic cat and comparing the error rate with that of untreated products. Significant reduction of all mutation types was observed. Statistical analysis revealed that the T4 and E.coli endonucleases reduced the occurrence of mutations in cloned synthetic gene products. The EMC treatment was successful especially in the removal of deletions and insertions from the primary ligation products.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.