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.
Reddy, T. R., Kelsall, E. J., Fevat, L. M., Munson, S. E., Cowley, S. M. Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors. J. Vis. Exp. (95), e52155, doi:10.3791/52155 (2015).