Eppendorf Multiporator - Manual (Page 17)

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4.7.2 Electroporation buffer for eukaryotic cells
To achieve optimal transfection results, the original electroporation buffer
from Eppendorf, with a low electrical conductivity, should be used. These
electroporation buffers (hypo and isoosmolar) are tested for sterility and
the absence of mycoplasms and endotoxins.
The use of these buffer systems means that considerably less current
flows during electroporation, with the result that no significant damage is
sustained by the cells. Simultaneously, the use of appropriate buffer
media results in the electrically induced "pores" being much larger than
those of pulse applications in conductive solutions.
Ideally, the electroporation should be carried out in hypoosmolar buffer.
Through the hypoosmolar buffer system, the cells absorb water shortly
before the pulse and swell as a result. Due to a variety of effects, including
the lowering of the breakdown voltage, this process enables increased
permeability of the plasma membrane. The yields can thereby be
considerably increased in comparison to those under isoosmolar
conditions. In the case of cells that react sensitively to purely hypoosmolar
conditions, step-by-step addition of the isoosmolar electroporation buffer
can be used to adjust the necessary osmolarity.
The Eppendorf electroporation buffer should be used for the electro-
poration of eukaryotic cells (with the exception of yeasts and some
microorganisms and plants). Electroporation media with low conductivity
can be used for the electroporation of bacteria, yeasts and other
microorganisms. Detailed application protocols can be downloaded from
the Eppendorf Homepage www.eppendorf.com.
4 Device description
Multiporator_Text_en.fm Seite 59 Freitag, 9. Juni 2006 8:05 08