6 factors affecting electroporation, Factors affecting electroporation, Chapter 6. factors affecting electroporation – Bio-Rad Gene Pulser MXcell™ Electroporation System User Manual
Page 50: Waveforms
Gene Pulser MXcell™ System Manual
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6
Factors Affecting
Electroporation
The Gene Pulser MXcell™ electroporation system can help you design successful
electroporation experiments. Read the sections in this chapter for information that can assist
you in designing your electroporation experiments:
•
Factors affecting electroporation (page 41)
•
Electroporation Theory (page 45)
Factors Affecting Electroporation
When considering electroporation of an unfamiliar cell line, we generally recommend
reviewing the protocols from several references and making a consensus starting protocol. If
no references exist for your particular cell line of interest, then we suggest using references
for a similar cell type as a starting point. Alternatively, select the Opt mini 96-well/ Sqr, Exp
pre-set protocol (page 34) to find initial conditions, then fine tune the conditions.
The following sections discuss different factors that affect the success of an electroporation
experiment:
•
Waveform (page 41)
•
Duration and number of pulses (page 42)
•
Cell growth (page 42)
•
Nucleic acids and biomolecules (page 42)
•
Electroporation buffer (page 43)
•
Temperature (page 44)
Waveforms
The two most common waveforms used in electroporation today are the square wave and
exponential. The square wave relies on a charge being applied to the cells for a set time. The
exponential waveform builds up a charge in a capacitor, and when applied to the sample,
the voltage delivered decays exponentially, until the charge remaining is about 37% of the
original pulse. The time over which this decay occurs is known as the time constant and is
equal to (RxC) where the resistance of the sample and system is R and the capacitance set
on the instrument is C.