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Patched circle PCR
A method for site-directed deletion of a large-scale fragment from a cloning vector, e.g. for obtaining cDNA from a plasmid without vector sequences. Two PCR primers pair to the 5'- and 3'-flanking sequences adjacent to and extending away from the unwanted sequence, and the DNA is amplified by PCR. Another oligonucleotide is added to bind to both ends of the amplified linear cDNA and patches them together to mimic closed circular DNA, which is then used to transform E. coli.
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- Category: Biochemistry; Molecular biology
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