SMC, a simple method to rapidly assemble multiple fragments into one construct.
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.
Tan
Guihong
G
Division of Biological Sciences, Bond Life Sciences Center, University of Missouri, MO 65211, USA. tanc@missouri.edu
Tan
Change
C
D001483:Base Sequence; D017931:DNA Primers; D004271:DNA, Fungal / Q000737:chemistry*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015003:Yeasts / Q000737:chemistry*
D017931:DNA Primers; D004271:DNA, Fungal