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PMID: 4000951 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

Nucleic acids research ·Vol. 13 ·No. 6 ·1985-03-25 ·Pages 1997-2008

Rusche JR, Howard-Flanders P

Abstract

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.

MeSH Terms
Cations, Monovalent/pharmacology Chlorides Cobalt/pharmacology DNA/metabolism DNA Ligases/metabolism Kinetics Polyamines/pharmacology Polynucleotide Ligases/metabolism T-Phages/enzymology
Chemicals
Cations, Monovalent Chlorides Polyamines hexaminecobalt (III) trichloride Cobalt DNA DNA Ligases Polynucleotide Ligases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rusche J R
Howard-Flanders P
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20 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.6.1997
Published
1985-03-25
Pages
1997-2008
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341130
Subset
IM
Grants
NCI NIH HHS · CA 09259-06 · United States
NCI NIH HHS · CA 20763 · United States
External Links
PubMed source DOI: 10.1093/nar/13.6.1997 PMC Full Text View journal details
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