Journal
BIOTECHNIQUES
Volume 43, Issue 3, Pages 356-359Publisher
FUTURE SCI LTD
DOI: 10.2144/000112536
Keywords
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Funding
- NIAID NIH HHS [AI54456, AI39671, AI56299] Funding Source: Medline
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In-Fusion (TM) can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, we have found In-Fusion can join four or more pieces of DNA in a single reaction. We used this insight to construct seamless fusion proteins, modular vectors with readily interchangeable segments, and novel mutagenesis strategies. Replacement In-Fusion can be used to delete any desired DNA segment in a plasmid and replace it with any desired new DNA segment without limitations on position or size.
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