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Nucleic Acids Research, 1977, Vol. 4, No. 1 85-98
© 1977

Articles

Equimolar addition of oligoribonucleotides with T4 RNA ligase

Olke C. Uhlenbeck and Vicki Cameron

Department of Biochemistry, University of Illinois IL 61801, USA

Received September 20, 1976. 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.


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