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Nucleic Acids Research, 1985, Vol. 13, No. 7 2241-2249
© 1985

Articles

Macromolecular crowding accelerates the cohesion of DNA fragments with complementary termini

Steven B. Zimmerman and Barbara Harrison

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

Received January 14, 1985. Revised March 8, 1985. Accepted March 8, 1985.

Macromolecular crowding Increases the rate of nonenzymatic cohesion of the complementary ends of {lambda} DNA. Both {lambda} DNA and DNA fragments bearing the cohesive ends of {lambda} DNA are similarly affected. High concentrations of plasma albumin or Ficoll 70 increase the rate of cohesion by ca. 100-fold whereas high concentrations of polyethylene glycol 8000 cause >2000-fold stimulation in this rate. These results have implications for the mechanism of polymer-stimulated enzymatic ligation of DNA or RNA. In addition, these crowding effects may help to explain the rapid cohesion of {lambda} DNA observed in vivo.

An improved procedure for the recovery of DNA fragments separated by agarose gel electrophoresis is also described.


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