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Team:Harvard/Lambda Red
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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 [https://2011.igem.org/Team:Harvard/Protocols#Lambda_Red Protocols] section. See [https://2011.igem.org/Team:Harvard/Results/One-Hybrid_Selection#Building_the_selection_strain:_Lambda_Red_Recombineering Lambda Red results] section for how we used lambda red to build our selection system. | 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 [https://2011.igem.org/Team:Harvard/Protocols#Lambda_Red Protocols] section. See [https://2011.igem.org/Team:Harvard/Results/One-Hybrid_Selection#Building_the_selection_strain:_Lambda_Red_Recombineering Lambda Red results] section for how we used lambda red to build our selection system. | ||
===Lambda red in ECNR2=== | ===Lambda red in ECNR2=== | ||
| + | We used a phage lambda-derived Red recombination system because of its efficiency | ||
| + | |||
===How to use ECNR2=== | ===How to use ECNR2=== | ||
For efficient genome editing using lambda red, you can use [https://2011.igem.org/Team:Harvard/Results/Biobricks#EcNR2_strain_.28BBa_K615002.29 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.] | For efficient genome editing using lambda red, you can use [https://2011.igem.org/Team:Harvard/Results/Biobricks#EcNR2_strain_.28BBa_K615002.29 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.] | ||
Revision as of 01:12, 29 September 2011
Overview | MAGE | Chip-Based Synthesis | Lambda Red | Protocols
Contents |
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
We used a phage lambda-derived Red recombination system because of its efficiency
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]
