Team:Harvard/Lambda Red

Lambda Red

Lambda red recombineering makes use of homologous recombination systems to allow the insertion of constructs into the genome. It is accomplished in two steps, as shown in the diagram below and in our Protocols section. See Lambda Red results section for how we used lambda red to build our selection system.

Lambda red in ECNR2

How to use ECNR2

For efficient genome editing using lambda red, you can use this strain with your own insertion construct and overhangs. To obtain this strain, you can make a request at [http://partsregistry.org/Part:BBa_K615002 the Registry of Standard Biological Parts.]

Figure 1. Lambda Red, PCR to get the required insertion product (zeocin in this example) 915px

References:

[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC165854/ Yu, D., H. M. Ellis, et al. (2000). "An efficient recombination system for chromosome engineering in Escherichia coli." Proceedings of the National Academy of Sciences of the United States of America 97(11): 5978-5983.]

[http://www.genetics.org/content/186/3/791 Mosberg JA, Lajoie MJ, Church GM. Lambda red recombineering in Escherichia coli occurs through a fully single-stranded intermediate. Genetics 2010;186:791-799.]

[http://www.sciencemag.org/content/suppl/2011/07/13/333.6040.348.DC1/Isaacs.SOM.pdf ECNR2 strain details]