Seamless cloning and domain swapping of synthetic and complex DNA.
Anal Biochem
; 385(1): 171-3, 2009 Feb 01.
Article
em En
| MEDLINE
| ID: mdl-18976627
ABSTRACT
The use of synthetic DNA can avoid problems that are sometimes encountered with conventional molecular biology techniques using DNA with high GC content, strong secondary structures, or repeat sequences. However, very complex DNA may still resist PCR and synthesis of DNA from oligonucleotides. In the method described here, separately synthesized DNA segments were seamlessly joined independently of the presence of restriction sites in the target DNA. This method allowed the reconstruction of complex DNA by concatenation of easily synthesized segments and permitted repeated swapping of segments, from a few nucleotides to large fragments of complex DNA for phenotypic analysis.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Clonagem Molecular
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2009
Tipo de documento:
Article