Rapid hierarchical assembly of medium-size DNA cassettes.
Nucleic Acids Res
; 40(12): e92, 2012 Jul.
Article
in En
| MEDLINE
| ID: mdl-22422837
ABSTRACT
Synthetic biology applications call for efficient methods to generate large gene cassettes that encode complex gene circuits in order to avoid simultaneous delivery of multiple plasmids encoding individual genes. Multiple methods have been proposed to achieve this goal. Here, we describe a novel protocol that allows one-step cloning of up to four gene-size DNA fragments, followed by a second assembly of these concatenated sequences into large circular DNA. The protocols described here comprise a simple, cheap and fast solution for routine construction of cassettes with up to 10 gene-size components.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cloning, Molecular
Language:
En
Journal:
Nucleic Acids Res
Year:
2012
Type:
Article
Affiliation country:
United States