Y-ligation: an efficient method for ligating single-stranded DNAs and RNAs with T4 RNA ligase.
Very efficient ligation of oligodeoxyribonucleotides was attained through a simple molecular construct, which is composed of one stem and two branches (Y-shape), with use of T4 RNA ligase. Single-stranded DNAs (naturally, RNAs also) of more than 100 nucleotides (even 800 nts) were considerably ligated, approximately as theoretically expected. Owing to the molecular construct adopted, such a tiny amount of ligation products could be amplified to a sufficient amount by PCR and then recovered as single-stranded DNAs. This advantage of being amplifiable is shown to be useful for both combinatorial chemistry and evolutionary molecular engineering, which deal with a pool of diversity molecules.
Nishigaki
K
K
Department of Functional Materials Science, Saitama University, Urawa, Japan. koichi@fms.saitama-u.ac.jp
Taguchi
K
K
Kinoshita
Y
Y
Aita
T
T
Husimi
Y
Y
D001698:Biomedical Engineering; D020650:Combinatorial Chemistry Techniques; D004277:DNA, Single-Stranded / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000378:metabolism*; D011130:RNA Ligase (ATP) / Q000378:metabolism*
D004277:DNA, Single-Stranded; D012313:RNA; D011130:RNA Ligase (ATP)