Equimolar addition of oligoribonucleotides with T4 RNA ligase (original) (raw)

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Department of Biochemistry, University of Illinois

IL 61801, USA

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Department of Biochemistry, University of Illinois

IL 61801, USA

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Received:

20 September 1976

Published:

01 January 1977

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Olke C. Uhlenbeck, Vicki Cameron, Equimolar addition of oligoribonucleotides with T4 RNA ligase, Nucleic Acids Research, Volume 4, Issue 1, 1 January 1977, Pages 85–98, https://doi.org/10.1093/nar/4.1.85
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Abstract

T4 induced RNA ligase will join equimolar concentrations of two oligo-ribonucleotides, (Ap) 3 C and p(Up) 5 , to form a single product, (Ap) 3 Cp(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.

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