Gateway Recombinational Cloning
Program in Systems Biology; Program in Molecular Medicine
Genetic Phenomena | Genetics and Genomics | Genetic Structures | Investigative Techniques | Laboratory and Basic Science Research | Molecular Biology | Systems Biology
The Gateway recombinatorial cloning system was developed for cloning multiple DNA fragments in parallel (e.g., in 96-well formats) in a standardized manner using the same enzymes. Gateway cloning is based on the highly specific integration and excision reactions of bacteriophage lambda into and out of the Escherichia coli genome. Because the sites of recombination ("att" sites) are much longer (25-242 bp) than restriction sites, they are extremely unlikely to occur by chance in DNA fragments. Therefore, the same recombination enzyme can be used to robustly clone many different fragments of variable size in parallel reactions.
DOI of Published Version
Cold Spring Harb Protoc. 2018 Jan 2;2018(1):pdb.top094912. doi: 10.1101/pdb.top094912. Link to article on publisher's site
Cold Spring Harbor protocols
Reece-Hoyes JS, Walhout AJ. (2018). Gateway Recombinational Cloning. Program in Systems Biology Publications. https://doi.org/10.1101/pdb.top094912. Retrieved from https://escholarship.umassmed.edu/sysbio_pubs/119