Fusion of restriction termini using synthetic adaptor oligonucleotides.
A method is proposed for linking a DNA fragment possessing a 5' single-stranded extension to one carrying a 3' extension. Synthetic oligonucleotide adaptors can be used to (i) change the site specificity at the termini of a fragment generated by restriction enzyme cleavage and (ii) simultaneously dephosphorylate the extremities of a DNA molecule to prevent recircularisation and allow positive selection for recombinant DNA molecules.
D000595:Amino Acid Sequence; D001483:Base Sequence; D011088:DNA Ligases; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics*; D010957:Plasmids; D013604:T-Phages / Q000201:enzymology
D004274:DNA, Recombinant; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D004262:DNA Restriction Enzymes; D011088:DNA Ligases