Protocols

Our lab has published several protocols on recombineering. We strongly encourage you to consult several of them. Depending on your exact needs, parts of protocols will need to be mixed and matched. At the bottom of this page, you can also download some more general protocols we use in the lab.

  •     Thomason, L. C., Sawitzke, J. A., Li, X., Costantino, N. & Court, D. L. (2014). Recombineering: genetic engineering in bacteria using homologous recombination. Curr Protoc Mol Biol 106, 1 16 1-1 16 39. CPMB 2014.pdf.

        Sawitzke, J. A., Thomason, L. C., Costantino, N., Bubunenko, M., Datta, S. & Court, D. L. (2007). Recombineering: in vivo genetic engineering in E. coli, S. enterica, and beyond. Methods Enzymol. 421, 171-199. MIE.pdf 

  •     Sawitzke, J. A., Costantino, N., Li, X. T., Thomason, L. C., Bubunenko, M., Court, C. & Court, D. L. (2011). Probing Cellular Processes with Oligo-Mediated Recombination and Using the Knowledge Gained to Optimize Recombineering. J. Mol. Biol. Oligo optimization.pdf

  •     Sharan, S. K., Thomason, L. C., Kuznetsov, S. G. & Court, D. L. (2009). Recombineering: a homologous recombination-based method of genetic engineering. Nat. Prot. 4, 206-223. BACS.pdf

        Chan, W., Costantino, N., Li, R., Lee, S.C., Su, Q., Melvin, D., Court, D.L., and Liu, P. (2007). A recombineering based approach for high-throughput conditional knockout targeting vector construction. Nucleic Acids Res 35, e64. high throughput.pdf

  •     Thomason, L. C., Costantino, N., Shaw, D. V. & Court, D. L. (2007). Multicopy plasmid modification with phage  Red recombineering. Plasmid 58, 148-158. plasmids.pdf

  •     Thomason, L. C., Oppenheim, A. B. & Court, D. L. (2009). Modifying bacteriophage lambda with recombineering. Methods Mol Biol 501, 239-51.phage.pdf

    We also recommend the following reference for an excellent method for recombineering phage: 

            Marinelli, L. J., Piuri, M., Swigonova, Z., Balachandran, A., Oldfield, L. M., van Kessel, J. C. & Hatfull, G. F. (2008). BRED: a simple and powerful tool for constructing mutant and recombinant bacteriophage genomes. PLoS ONE 3, e3957.