IMR Press / FBE / Volume 2 / Issue 3 / DOI: 10.2741/E169

Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on as a courtesy and upon agreement with Frontiers in Bioscience.

SMC, a simple method to rapidly assemble multiple fragments into one construct
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1 Division of Biological Sciences, University of Missouri-Columbia, MO 65211, USA
Front. Biosci. (Elite Ed) 2010, 2(3), 1105–1114;
Published: 1 June 2010

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 ~30 base pair overlaps in yeast using raw PCR and/or restrict enzyme-digested products. We assembled seven DNA fragments into one contig in a single step by SMC in eight days. 

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