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Protocols for C-Brick DNA Standard Assembly Using Cpf1
Published on: June 15, 2017
BioBrick™ assembly using the In-Fusion PCR Cloning Kit
Sean C Sleight1, Herbert M Sauro
1Department of Bioengineering, University of Washington, Seattle, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|September 3, 2013
Summary
Synthetic biologists can now assemble genetic circuits using the In-Fusion PCR Cloning Kit, an alternative to traditional methods. This new protocol offers a faster and more efficient way to combine standardized biological parts, known as BioBricks™.
Area of Science:
- Synthetic Biology
- Molecular Biology
- Genetic Engineering
Background:
- Synthetic biology relies on assembling standardized genetic parts called BioBricks™.
- Traditional BioBrick™ assembly uses restriction enzymes and DNA ligase, often a stepwise process.
- Efficient assembly methods are crucial for constructing complex genetic circuits.
Purpose of the Study:
- To describe an alternative protocol for assembling two BioBricks™.
- To demonstrate the utility of the In-Fusion PCR Cloning Kit for BioBrick™ assembly.
- To present a protocol adaptable to other recombination-based assembly methods.
Main Methods:
- Utilized the In-Fusion PCR Cloning Kit for assembling two BioBricks™.
- Described a protocol adaptable to recombination-based methods like SLIC and Gibson assembly.
- Focused on the efficient joining of standardized DNA parts.
Main Results:
- Successfully demonstrated BioBrick™ assembly using the In-Fusion PCR Cloning Kit.
- Provided a protocol that can be adapted for various recombination-based DNA assembly techniques.
- Offered an alternative to conventional restriction enzyme-based assembly.
Conclusions:
- The In-Fusion PCR Cloning Kit provides an effective alternative for BioBrick™ assembly.
- This protocol facilitates the construction of genetic circuits by streamlining the assembly of biological parts.
- Adaptability to other recombination methods broadens its applicability in synthetic biology research.
