The gene pulser mxcell electroporation guide – Bio-Rad Gene Pulser MXcell™ Electroporation System User Manual
Page 2
To choose a starting
set of protocols,
search our library of
electroprotocols at
or search
The Gene Pulser MXcell electroporation system is filled with features that will enable
you to quickly optimize conditions for efficient delivery of molecules into most eukaryotic
cells, including mammalian cells and plant protoplasts. This guide will assist you in
determining the parameters for obtaining efficient transfection of molecules into your
favorite cells while maintaining high cell viability.
Transfection efficiency and cell viability
Transfection and cell viability are affected by:
n
Cell type and its physiological condition and state prior to electroporation:
cells should be actively growing, healthy, and free of contamination
n
Temperature of buffer during electroporation: when using Gene Pulser
®
electroporation
buffer, electroporation should be performed at room temperature
n
Cell density: for most experiments using the 96-well plate, we recommend using
150 µl of cells/well at a density of 1 x 10
6
to x 10
6
cells/ml in Gene Pulser
electroporation buffer
n
Concentration of the transfected molecule (RNA, DNA): ideally a concentration
of 5–40 µg/ml should be used for plasmid DNA and 10–100 nM for siRNA (only
Bio-Rad’s siLentMer
™
Dicer-substrate siRNA duplexes)
n
The characteristics of the electric pulse
Electroporation protocols
Electroporation applies an electric pulse to cells to promote uptake of molecules (RNA,
DNA) into the cells. The electric pulse is defined by parameters that can be programmed
as protocols in the Gene Pulser MXcell. Two type of pulses can be delivered by the
Gene Pulser MXcell:
the exponential waveform which is defined by voltage,
capacitance, and resistance or the square waveform which is defined by voltage,
pulse duration, and resistance.
The Gene Pulser MXcell can be programmed three ways:
n
Manually, under “Protocol Setup”
n
By creating a voltage gradient under “Gradient Protocol”
n
By using preprogrammed protocols under “Pre-Set Protocols”*
This guide will help you program the Gene Pulser MXcell whether you have prior
knowledge of optimal electroporation conditions for your cells or not.
Gene Pulser MXcell
Electroporation Guide
The Gene Pulser MXcell
Electroporation Guide
| Electroporation Optimization Overview
* Note: The parameters
entered into the preset
protocols are based
on proven conditions
used for electroporation
of mammalian cells and
take advantage of the
Gene Pulser MXcell’s
ability to apply multiple
conditions simultaneously.
The preset protocols
can be edited and
stored in your user
file for future use.