One-pot fusion polymerase chain reaction for combinatorial synthesis of DNA from several cassettes.
The fusion of DNA fragments is becoming increasingly more important. The ability to work without being constrained by restriction sites enables DNA fusion to be applied to a much broader range of fragments. Therefore, we describe a simplified polymerase chain reaction (PCR)-based method for fusion of DNA fragments in one step. In a single PCR, two templates, an overlapping primer, and template-specific forward and reverse primers are used. After a few cycles, the fusion DNA is assembled and is amplified. The ratio of overlapping primer to forward/reverse primers and template DNA is essential for the success of the reaction.
Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, D-38106 Braunschweig, Germany.
Malte
M
Büttner
Leif
L
Barleben
D019677:Artificial Gene Fusion / Q000379:methods*; D004247:DNA / Q000096:biosynthesis*; D017343:Genes, Plant; D016133:Polymerase Chain Reaction / Q000379:methods*
D004247:DNA