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Updated: Feb 1, 2026
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Studying DNA Looping by Single-Molecule FRET
Published on: June 28, 2014
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Loop assembly: a simple and open system for recursive fabrication of DNA circuits
Bernardo Pollak1, Ariel Cerda2,3, Mihails Delmans1
1Department of Plant Sciences, University of Cambridge, Cambridge, CB2 3EA, UK.
The New Phytologist
|December 7, 2018
Summary
Loop assembly offers a recursive DNA fabrication method for creating large, complex synthetic DNA circuits. This efficient system simplifies DNA construction for applications like plant transformation.
Area of Science:
- Synthetic Biology
- Molecular Biology
- Biotechnology
Background:
- Existing DNA assembly methods require customization and complex procedures.
- There is a need for efficient and versatile DNA fabrication techniques.
Purpose of the Study:
- To develop a recursive DNA fabrication system for efficient assembly of large DNA circuits.
- To provide a generalized solution for DNA construction with standardized parts.
Main Methods:
- Developed Loop assembly, a recursive DNA fabrication system utilizing two Type IIS restriction endonucleases and corresponding vector sets.
- Assembled standardized level 0 parts into circuits of 1, 4, 16 or more genes by looping between two vector sets.
- Incorporated modular sites for hybrid assembly using sequence overlap methods.
Main Results:
- Successfully constructed plasmids up to 16 genes and 38 kb with high efficiency (>80%).
- Characterized Loop assembly on over 200 DNA constructs.
- Validated method fidelity using high-throughput Illumina plasmid sequencing.
Conclusions:
- Loop assembly provides an efficient and versatile DNA fabrication method.
- The system simplifies DNA construction for applications such as plant transformation.
- The cloning system is available under an OpenMTA license for open access.
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