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Oligonucleotide fragments can be directionally subcloned into vectors at a single restriction site. By using T4 DNA polymerase exonuclease activity to treat vector DNA, single-stranded ends can be generated. The oligonucleotide sequences are designed to have sequence complementary to these single-stranded ends. Through the homologous annealing of oligonucleotides to the treated vector ends, the...
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