Ligation with hammerhead ribozymes
OwnerDavid M. Long, Olke C. Uhlenbeck +1inventors
InventorsDavid M. Long, Olke C. Uhlenbeck, Klemens J. Hertel
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Abstract
The claimed invention is drawn to a method for ligation of an enzymatic nucleic acid molecule having a hammerhead motif to a separate nucleic acid molecule. The method includes use of ligation enhancing conditions in which ligation is enhanced at least two fold over that observed in a medium containing only Mg.sup.2+ as a divalent cation at 20.degree. C.
Claims15
A method for ligation of an enzymatic nucleic acid molecule having a hammerhead motif to an RNA molecule, comprising the steps of:
- contacting in a reaction mixture said enzymatic nucleic acid molecule with said RNA molecule wherein said reaction mixture contains an ion selected from the group consisting of:
- cobalt, manganese, cadmium, zinc, copper, and nickel ions, in which ligation is enhanced at least two fold over that observed in a medium containing only Mg.sup.2+ as a divalent cation at 20.degree. C.
A method for generating efficient ligation of a hammerhead ribozyme to a target RNA comprising ligation condition comprising manganese or cobalt ions at a temperature below 20.degree. C.
A method for selection of a ligation efficient enzymatic nucleic acid molecule with a hammerhead motif, comprising the steps of selecting for hammerhead molecules able to ligate to separate RNA molecules under ligation enhancing conditions wherein said ligation enhancing conditions comprise an ion selected from the group consisting of:
- cobalt, manganese, cadmium, zinc, copper, and nickel ions.
Dependencies are inferred from the claim text.