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Team:Harvard/Lambda Red
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==References:== | ==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.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/early/2010/09/02/genetics.110.120782.full.pdf Mosberg JA, Lajoie MJ, Church GM. Lambda red recombineering in Escherichia coli occurs through a fully single-stranded intermediate. Genetics 2010;186:791-799.] | ||
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Revision as of 15:06, 26 September 2011
Overview | MAGE | Chip-Based Synthesis | Lambda Red | Protocols
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. Also see Lambda Red results for how we used lambda red to build our selection system.
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/early/2010/09/02/genetics.110.120782.full.pdf Mosberg JA, Lajoie MJ, Church GM. Lambda red recombineering in Escherichia coli occurs through a fully single-stranded intermediate. Genetics 2010;186:791-799.]
