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Nucleic Acids Res. 1983 November 25; 11(22): 7853–7871.
PMCID: PMC326544

Polymer-stimulated ligation: enhanced blunt- or cohesive-end ligation of DNA or deoxyribooligonucleotides by T4 DNA ligase in polymer solutions.

Abstract

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Zimmerman SB, Pheiffer BH. Macromolecular crowding allows blunt-end ligation by DNA ligases from rat liver or Escherichia coli. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5852–5856. [PubMed]
  • Gellert M, Mizuuchi K, O'Dea MH, Nash HA. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. [PubMed]
  • McDonell MW, Simon MN, Studier FW. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. [PubMed]
  • Maniatis T, Jeffrey A, van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. [PubMed]
  • Sugino A, Goodman HM, Heyneker HL, Shine J, Boyer HW, Cozzarelli NR. Interaction of bacteriophage T4 RNA and DNA ligases in joining of duplex DNA at base-paired ends. J Biol Chem. 1977 Jun 10;252(11):3987–3994. [PubMed]
  • Krasnow MA, Cozzarelli NR. Catenation of DNA rings by topoisomerases. Mechanism of control by spermidine. J Biol Chem. 1982 Mar 10;257(5):2687–2693. [PubMed]
  • Deugau KV, van de Sande JH. T4 polynucleotide ligase catalyzed joining of short synthetic DNA duplexes at base-paired ends. Biochemistry. 1978 Feb 21;17(4):723–729. [PubMed]
  • Roberts RJ, Wilson GA, Young FE. Recognition sequence of specific endonuclease BamH.I from Bacillus amyloliquefaciens H. Nature. 1977 Jan 6;265(5589):82–84. [PubMed]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press