Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol Protocol describes transfection of plasmid DNA into mammaliancell lines using electroporation, a process whereby externalapplication of electric pulses induce cell membrane permeability.A number of factors can affect electrotransfection efficiency.In general, cells in suspension and small volume cells are difficultto transfect, whereas adherent cells and large volume cellsare relatively easy.
Protocol Electroporation Protocol describes a method for introducing gene constructsinto mouse embryos by electroporation. Gene constructs can bequickly tested for tissue-specific transcriptional activityor can be used to overexpress gene products.
Electrocompetent bacteria are prepared by growing cultures tomid-log phase, washing the bacteria extensively at low temperature,and then resuspending them in a solution of low ionic strengthcontaining glycerol. DNA is introduced during exposure of thebacteria to a short high-voltage electrical discharge.
Transformation of Tetrahymena thermophila by Electroporation Protocol Protocol describes a method for transformation of the Tetrahymenausing electroporation. The vector is electroporated into cellsafter mating, where it is incorporated into the DNA of developingmacronuclei. Because T. thermophila can be propagated indefinitelywithout conjugation, transformation of the macronucleus providesa way to obtain stable somatic transformants.