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Nucleic Acids Research, 1984, Vol. 12, No. 4 1791-1810
© 1984

MOLECULAR BIOLOGY

Fluorescent labelling of tRNA and oligodeoxynucleotides using T4 RNA ligase

Richard Cosstick, Larry W. McLaughlin and Fritz Eckstein

Max-Planck-Institut für experimentelle Medizin, Abteilung Chemie Hermann-Rein-Strasse 3, D-3400 Göttingen, FRG

Received December 19, 1983. Accepted January 18, 1984.

3'-O-(5'-phosphoryldeoxycytidyl) phosphorothioate and fluorescent 3'-O-(5'-phosphoryldeoxycytidyl) S-bimane phosphorothioate can be ligated to tRNA by T4 RNA ligase. They are also efficient donors for the enzymatic ligation to oligodeoxynucleotides bearing a 3'-cytidine terminus. Cytidine 3', 5'-bisphosphate is also a substrate for the ligation reaction with DNA restriction fragments with a 3'-terminate cytidylic acid residue. Oligo- and poly nucleotides with a 3' -phosphorothiote group react readily with electrophiles as exemplified by the reaction with monobromobimane.


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