Eppendorf Multiporator - Electroporation - Manual (Page 4)

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The phenomenon whereby a short, intensive current surge (pulse) is used to generate reversible openings (pores) in a
membrane was first used in the 1970s to introduce foreign molecules into cells. These membrane pores allow low-
molecular substances (such as dyes or peptides) and high-molecular substances (such as proteins, DNA and RNA) to be
introduced into cells. This procedure, which is known as electroporation, electroinjection or electropermeabilization, has
developed into a standard method in many laboratories, and is used primarily for the transfection of eukaryotic cells and
bacteria.
The Eppendorf Multiporator ® and the special electroporation buffers form a system which allows electroporation to take
place efficiently and gently under hypoosmolar conditions. The hypoosmolar buffer causes the cells to swell up, which
expands the membrane and loosens up the cytoskeleton. This in turn leads to a reduction in voltage required for the
formation of membrane pores. Electroporation can thus be performed in a more "cell-friendly" manner without any
adverse effect on transfection efficiency.
This guide is valid for the eukaryotic module of the Multiporator, by which eukaryotic cells (except yeast and some
microorganisms) can be electroporated.
It contains concise descriptions of the experimental conditions that form the basis of the innovative electroporation
process carried out using the Multiporator's Soft Pulse technology. Time should be taken to familiarize oneself with the
effects that important parameters of the Soft Pulse technology, such as pulse voltage, time, temperature and the
composition of the medium, have on the transfection yield. This will enable the best possible transfection results to be
achieved with a specific cell line.
Any experiences with commonly used electroporation techniques in the past utilizing millisecond pulse times cannot be
compared directly to the technology using the Multiporator
® with microsecond pulse times.
Application protocols for many different cell lines can be found on the Eppendorf homepage at www.eppendorf.com.
If cells are used for which no application protocol is available, Section 3, "Optimizing the parameters", and the general
electroporation protocol in Section 4 should be consulted. Section 5, "Troubleshooting", contains assistance for those
occasions when results have not turned out as expected.
Bacteria, yeast as well as other microorganisms can be transformed with the optional bacteria module. Application
protocols are available at www.eppendorf.com too.
1.1 Purpose of the applications guide
1 Introduction
1 Introduction
Multipor_Appli_E_poration_en.fm Seite 28 Montag, 30. Januar 2006 2:17 14