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Nucleic Acids Research, 1983, Vol. 11, No. 22 7853-7871
© 1983

MOLECULAR BIOLOGY

Polymer-stimulated ligation: enhanced blunt- Or cohesive-end ligation of DNA or deoxyribooligonudcleotides by T4 DNA Ugase in polymer solutions

Barbara H. Pheiffer and Steven B. Zimmerman

Laboratory of Molecular Biology, National Institute of Arthritis, Diabetes and Digestive and Kidney Diseases, National Institutes of Health Bethesda, MD 20205,USA

Received August 23, 1983. Revised October 19, 1983. Accepted October 19, 1983.

The rates of blunt-end and cohesive-end ligation of DNA by T4 DNA ligase are Increased by orders of magnitude in the presence of high concentrations of a variety of nonspecific polymers such as polyethylene glycol, Ficoll, bovine plasma albumin, or glycogen. Blunt-end ligation of small self-complementary oligodeoxyribonucleotides is also stimulated. The use of polyethylene glycol 6000 In such systems is characterized In some detail. Conditions are suggested which either greatly increase the rate of formation of and size of linear ligation products or which allow smaller but significant Improvements in the amounts of circular ligation products.


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