Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Calcium-Phosphate-Mediated Transfection of Eukaryotic Cells with Plasmid DNAs

(Protocol summary only for purposes of this preview site)

This protocol, which describes a calcium-phosphate-mediated transfection method for use with plasmid DNAs and adherent cells, was modified from Jordan et al. (1996), who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and human embryonic kidney 293 cells. Following this protocol are variations on the basic method.

  • For high-efficiency generation of stable transfectants, see the alternative method at the end of this protocol.
  • For use with high-molecular-weight genomic DNAs, see Protocol 3, and for use with adherent cells that have been released from the substratum with trypsin, see the alternative method at the end of Protocol 3.
  • For use with non-adherent cells, see the alternative method at the end of Protocol 3.


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