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Journal of tissue culture methods

, Volume 15, Issue 4, pp 176–180 | Cite as

High efficiency transfection of monkey kidney COS-1 cells

  • Douglas Mortlock
  • Elizabeth B. Keller
  • Cynthia J. Ziegra
  • Maja M. Suter
Article

Summary

Methods are described for achieving high efficiency transient transfection of COS-1 cells. A transfection solution of vector DNA and DEAE-dextran in phosphate buffered saline is added to the cells, followed by treatment with culture medium containing chloroquine, resulting in a maximum efficiency of about 50%. The high efficiency obtained is primarily dependent on the use of Ca2+- or Mg2+-free buffer solutions for transfection, because the addition of these ions greatly reduces efficiency. Shocking the cells with dimethylsulfoxide does not increase transfection efficiency. COS-1 cells are monkey kidney cells that have a genomic insertion of the SV40 T antigen gene, allowing plasmid expression vectors bearing an SV40 replication origin to be amplified in these cells. Therefore this transfection method is useful for optimizing transient expression of genes in a mammalian cell system.

Key words

DEAE-dextran transfection COS cells transient expression 

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Copyright information

© Tissue Culture Association 1993

Authors and Affiliations

  • Douglas Mortlock
    • 1
  • Elizabeth B. Keller
    • 1
  • Cynthia J. Ziegra
    • 1
  • Maja M. Suter
    • 1
  1. 1.Department of Pathology, College of Veterinary Medicine, and Section of Biochemistry, Molecular and Cell BiologyCornell UniversityIthaca

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