BASIC: A Simple and Accurate Modular DNA Assembly Method.
Biopart Assembly Standard for Idempotent Cloning (BASIC) is a simple, accurate, and robust DNA assembly method. The method is based on linker-mediated DNA assembly and provides highly accurate DNA assembly with 99 % correct assemblies for four parts and 90 % correct assemblies for seven parts [1]. The BASIC standard defines a single entry vector for all parts flanked by the same prefix and suffix sequences and its idempotent nature means that the assembled construct is returned in the same format. Once a part has been adapted into the BASIC format it can be placed at any position within a BASIC assembly without the need for reformatting. This allows laboratories to grow comprehensive and universal part libraries and to share them efficiently. The modularity within the BASIC framework is further extended by the possibility of encoding ribosomal binding sites (RBS) and peptide linker sequences directly on the linkers used for assembly. This makes BASIC a highly versatile library construction method for combinatorial part assembly including the construction of promoter, RBS, gene variant, and protein-tag libraries. In comparison with other DNA assembly standards and methods, BASIC offers a simple robust protocol; it relies on a single entry vector, provides for easy hierarchical assembly, and is highly accurate for up to seven parts per assembly round [2].
Department of Life Sciences, Centre for Synthetic Biology and Innovation, Imperial College London, London, SW7 2AZ, UK.
Marko
M
Storch
Department of Life Sciences, Centre for Synthetic Biology and Innovation, Imperial College London, London, SW7 2AZ, UK.
Arturo
A
Casini
Department of Life Sciences, Centre for Synthetic Biology and Innovation, Imperial College London, London, SW7 2AZ, UK.
Ben
B
Mackrow
Department of Bioengineering, Centre for Synthetic Biology and Innovation, Imperial College London, London, SW7 2AZ, UK. t.ellis@imperial.ac.uk.
Tom
T
Ellis
Department of Life Sciences, Centre for Synthetic Biology and Innovation, Imperial College London, London, SW7 2AZ, UK. g.baldwin@imperial.ac.uk.
Geoff S
GS
Baldwin
D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial
Combinatorial; DNA assembly; Hierarchical; Idempotent; Pathway engineering