Center for Fisheries, Environment and Aquaculture Sciences 1 article published in JoVE Biology Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors Thimma R. Reddy1, Emma J. Kelsall1, Léna M.S. Fevat2, Sarah E. Munson*3, Shaun M. Cowley*1 1Department of Biochemistry, University of Leicester, 2Center for Fisheries, Environment and Aquaculture Sciences, 3ES Cell Facility, Centre for Core Biotechnology Services, University of Leicester Gene targeting methodologies can be used to generate transgenic mice with knockout, knock-in and tagged alleles. Here, we describe an improved method of recombineering in E. coli, that we term ‘subcloning plus insertion’, which can be used to generate custom gene targeting vectors rapidly.
