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Homologous ligation
A method for fusion of two oligonucleotides, independently of any restriction site they share. A region flanking the desired ligation site on one fragment is introduced into the second fragment by PCR as part of one of the primers. The first fragment and the amplified second fragment are introduced in tandem into an appropriate plasmid that can be cleaved by a restriction nuclease engineered between the two fragments. The linearized DNA is used to transform a bacterium that has a high frequency of intramolecular homologous recombinations. Subsequent homologous recombination fuses the two fragments at their homologous region.
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- Category: Biochemistry; Molecular biology
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