A simple and highly efficient cloning method that employs PCR to directly create a fusion between insert and vector
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Updated: Aug 20, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
Published on: January 8, 2015
1Department of Life Science, National Tsing Hua University, Hsin-Chu 30055, Taiwan.
Abstract:
We have developed a very simple procedure for gene cloning using compound primers. This approach permits the insertion of a DNA fragment in a single step using a PCR-based cloning protocol, which employs annealing rather than ligation to create the recombinant in the plasmid. This method has been tested successfully to clone a Phalaenopsis gene coding for P450. A potential application of this protocol for constructing a cDNA library is also proposed.
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