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Nucleic Acids Research, 1985, Vol. 13, No. 6 1997-2008
© 1985

Articles

Hexamine cobalt chloride promotes intermolecular ligation of blunt end DNA fragments by T4 DNA ligase

James R. Rusche and Paul Howard-Flanders

Department of Therapeutic Rndiology, Yale University 260 Whitney Aye, 406 Gibbs, New Haven, CT 06511, USA

Received November 21, 1984. Revised February 27, 1985. Accepted February 27, 1985.

Hexamine cobalt chloride (HCC) increases the efficiency of blunt end ligation by T4 DNA ligase about 50 fold. Maximum stimulation occurs when standard buffers for ligation are supplemented with 1 mM HCC. All the ligation events are intermolecular regardless of the initial DNA concentration. In the presence of monovalent cations (eg. 25 mM KCl) HCC still increases the extent of T4 catalyzed ligation but intramolecular ligation products are also formed. Therefore, intermolecular ligation can be performed rapidly and at low DNA concentrations.


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