One-pot fusion polymerase chain reaction for combinatorial synthesis of DNA from several cassettes.

Malte Büttner, Leif Barleben

Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, D-38106 Braunschweig, Germany.

Analytical biochemistry 2012 Feb 15

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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. Copyright © 2011 Elsevier Inc. All rights reserved.

Citation

Malte Büttner, Leif Barleben. One-pot fusion polymerase chain reaction for combinatorial synthesis of DNA from several cassettes. Analytical biochemistry. 2012 Feb 15;421(2):797-8