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Nucleic Acids Research, 1985, Vol. 13, No. 22 7979-7992
© 1985

Articles

Stimulation of intermolecular ligation with E. coli DNA ligase by high concentration of monovalent cations in polyethylene glycol solutions

Ken'ichiro Hayashi, Masako Nakazawa, Yukuo Ishizaki, Nobutsugu Hiraoka and Akira Obayashi

Takara Shuzo Co. Ltd., Central Research Laboratories 3-4-1 Seta, Otsu-shi, Shiga 520-21, Japan

Received September 12, 1985. Revised October 31, 1985. Accepted October 31, 1985.

In the presence of high concentrations of the nonspecific polymer polyethylene glycol (PEG), intermolecular cohesive-end ligation with the DNA ligase from Escherichia coli was stimulated by high salt concentrations: 200 mM Nacl or 300 mM KCl in 10% (W/V) PEG 6000 solutions. Intermolecular blunt-end ligation with this ligase was also stimulated at 100–150 mM NaCl or 150–250 mM KCl in 15% PEG 6000 solutions. The extent of such intermolecular ligation increased and the salt concentrations at which ligation was stimulated extended to lower concentrations when we raised the temperature from 10 to 37°C.


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