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Volume 4, Issue 3, 1998, pp. 187-190

Y-ligation: An efficient method for ligating single-stranded DNAs and RNAs with T4 RNA ligase

Koichi Nishigaki, Katsuya Taguchi, Yasunori Kinoshita, Takuyo Aita, Yuzuru Husimi

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.

block shuffling, combinatorial chemistry, end-to-end distance, evolutionary molecular engineering, T4 RNA ligase, Y-ligation

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