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Synthetic double-stranded DNA oligonucleotides may be ligated into plasmid DNAs in order to introduce small sequence integrations/modifications. Briefly, the insert is not generated by PCR nor excised from an other vector but it is obtained by annealing two synthetic oligonucleotides. Oligos may be designed to have overhangs compatible with the termini generated by the restriction enzymes employed for linearizing the vector (check the short Addgene guide for further hints and information). Blunt-end cloning is possible but not advisable. The protocol includes three steps: oligonucleotide phosphorilation, oligonucleotide annealing and ds-oligo ligation into a linearized vector. Start from the second step (Annealing) if oligos were phosphorylated during synthesis Materials