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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 

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A new and efficient method for gene transfer into mouse FM3A cells using metaphase chromosomes by electroporation.

We introduced chromosome-mediated genes into mouse thymidine kinase-deficient FM3A (FM3Atk-) cells, by electroporation. The effects of some parameters on the electric shock-mediated transfection of FM3Atk- cells were investigated. Gene transfer of mouse L929 metaphase chromosome DNA into FM3Atk- resulted in a maximum frequency of (3.0 +/- 0.3) x 10(-5) at a cell density of 2.0 x 10(8)/ml and chromosome dosage of 5.0 x 10(7) cell equivalents/ml in a buffer containing 0.25 M mannitol, 0.5 mM MgCl2, 0.1mM CaCl2, and 1 mM Tris-HCl (pH 7.1). The highest yield of the transformants was obtained at an electric field strength of 1 kV/cm and a capacitance of 35 microF, with a single exponentially decaying pulse at 0 degrees C was optimal for post-shock incubation after electroporation. The tk gene was detected in the transformants by in situ hybridization analysis.[1]

References

  1. A new and efficient method for gene transfer into mouse FM3A cells using metaphase chromosomes by electroporation. Ohse, M., Tsuchida, K., Tomita, H., Taketo, A., Kimoto, H., Kusaoke, H. Biosci. Biotechnol. Biochem. (1996) [Pubmed]
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