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