Equimolar addition of oligoribonucleotides with T4 RNA ligase

Olke C. Uhlenbeck*, Vicki Cameron

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

77 Scopus citations

Abstract

T4 induced RNA ligase will join equimolar concentrations of two oligo-ribonucleotides, (Ap)3C and p(Up)5, to form a single product, (Ap)3Cp(Up) 5, in high yield. The presence of the 3′ phosphate on p(Up)5 prevents the oligomer from adding to itself. The pH optimum of the reaction is about 7.5, but less of the undesirable adenylated intermediate, App(Up)5, forms at pH 8.2. The reaction rate is a linear function of oligomer concentration from 3 μM to 0.6 mM. The data suggest that T4 RNA ligase will be a useful enzyme for the synthesis of oligomers of defined sequence.

Original languageEnglish (US)
Pages (from-to)85-98
Number of pages14
JournalNucleic acids research
Volume4
Issue number1
DOIs
StatePublished - Jan 1977

ASJC Scopus subject areas

  • Genetics

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