ArticleNucleic acids research2010
Isothermal DNA amplification using the T4 replisome: circular nicking endonuclease-dependent amplification and primase-based whole-genome amplification.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Thermostable in vitro transcription-translation compatible with microfluidic droplets.Microbial cell factories · 2024Article
- DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions.International journal of molecular sciences · 2023Review
- Isothermal Nucleic Acid Amplification to Detect Infection Caused by Parasites of the Trypanosomatidae Family: A Literature Review and Opinion on the Laboratory to Field Applicability.International journal of molecular sciences · 2022Review
- Article
- Towards a synthetic cell cycle.Nature communications · 2021Review
- Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology.Frontiers in bioengineering and biotechnology · 2020Review
- Toward long-lasting artificial cells that better mimic natural living cells.Emerging topics in life sciences · 2019Article
- Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System.Chembiochem : a European journal of chemical biology · 2019Article
- Self-replication of DNA by its encoded proteins in liposome-based synthetic cells.Nature communications · 2018Article
- The application of thermophilic DNA primase TtDnaG2 to DNA amplification.Scientific reports · 2017Article
- Article
- Linear nicking endonuclease-mediated strand-displacement DNA amplification.Analytical biochemistry · 2011Article
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6 authors.
Funding
Abstract
In vitro reconstitution of the bacteriophage T4 replication machinery provides a novel system for fast and processive isothermal DNA amplification. We have characterized this system in two formats: (i) in circular nicking endonuclease-dependent amplification (cNDA), the T4 replisome is supplemented with a nicking endonuclease (Nb.BbvCI) and a reverse primer to generate a well-defined uniform double-stranded linear product and to achieve up to 1100-fold linear amplification of a plasmid in 1 h. (ii) The T4 replisome with its primase (gp61) can also support priming and exponential amplification of genomic DNA in primase-based whole-genome amplification (T4 pWGA). Low amplification biases between 4.8 and 9.8 among eight loci for 0.3-10 ng template DNA suggest that this method is indeed suitable for uniform whole-genome amplification. Finally, the utility of the T4 replisome for isothermal DNA amplification is demonstrated in various applications, including incorporation of functional tags for DNA labeling and immobilization; template generation for in vitro transcription/translation and sequencing; and colony screening and DNA quantification.
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