Open Access
Article
SMC, a simple method to rapidly assemble multiple fragments into one construct
Guihong Tan1,Change Tan1
1
Division of Biological Sciences, Bond Life Sciences Center, University of Missouri, MO 65211, USA. tanc@missouri.edu
DOI: 10.2741/E169 Volume 2 Issue 3, pp.1105-1114
Published: 01 June 2010
Abstract

We describe a simple method, split-marker-mediated multiple-piece cloning (SMC), to rapidly assemble multiple DNA fragments into one construct in yeast. In this approach, a selectable marker is split into two non-functional, overlapping halves, of which one half is on the plasmid backbone. Homologous recombination reconstitutes the marker gene and assembles all DNA fragments in the desired order. This method allows rapid one-step fusion of various DNA fragments that contain approximately 30 base pair overlaps in yeast using raw PCR and/or restriction enzyme-digested products.

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Guihong Tan, Change Tan. SMC, a simple method to rapidly assemble multiple fragments into one construct. Frontiers in Bioscience-Elite. 2010. 2(3); 1105-1114.