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. 1993 Nov 25;21(23):5403-7.
doi: 10.1093/nar/21.23.5403.

Oligonucleotide circularization by template-directed chemical ligation

N G Dolinnaya  1 , M Blumenfeld, I N Merenkova, T S Oretskaya, N F Krynetskaya, M G Ivanovskaya, M Vasseur, Z A Shabarova
Affiliations
Free PMC article

Oligonucleotide circularization by template-directed chemical ligation

N G Dolinnaya et al. Nucleic Acids Res. .
Free PMC article

Abstract

An efficient method for producing the covalent closure of oligonucleotides on complementary templates by the action of BrCN was developed. A rational design of linear precursor oligonucleotides was studied, and the effect of factors such as oligonucleotide concentration and oligomer-template length ratio was evaluated. The efficiency of circularization was shown to correlate well with the secondary structure of the precursor oligomer (as predicted by a simple computer analysis), hairpin-like structures bearing free termini clearly favouring the circularization reaction. A novel idea, consisting of the incorporation of non-nucleotide insertions in the precursor oligomer (namely, 1,2-dideoxy-D-ribofuranose residues), may render this method universal and highly effective. An original set of assays was developed to confirm the circular structure of the covalently closed oligonucleotides.

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