Bio-Rad Gene Pulser MXcell Electroporation System User Manual

Gene Transfer

Gene Pulser MXcell

Electroporation Guide

™

To choose a starting

set of protocols,

search our library of

electroprotocols at

www.bio-rad.com/

genetransferprotocols/

http://www.ncbi.nlm.

* Note: The parameters

entered into the preset

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

or search

nih.gov/PubMed/.

protocols are based

can be edited and

stored in your user

file for future use.

Gene Pulser MXcell Electroporation Guide

The Gene Pulser MXcell
Electroporation Guide

The Gene Pulser MXcell electroporation system is filled with features that will enable
you to quickly optimize conditions for efficient deliver y 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 106 to 2 x 106 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 exponent ial 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.

2

Visit us on the Web at discover.bio-rad.com | Electroporation Optimization Overview

Schematic of 96-well
electroporation plate

One well set

Gene Pulser MXcell Electroporation Guide

Glossary

Electroporation plate: a plate in which electroporation is performed; available in

12-, 24-, and 96-well formats

Mini protocol: a preprogrammed set of 4–6 different protocols delivered to 4–6 well sets

Parameter s: the physical constants (waveform, voltage, capacitance, duration,

resistance) that define the electric pulse

Preset protocols: a set of preprogrammed protocols designed for rapid screening of

multiple parameters that can be used as a template

Protocol/electroporation conditions: parameters defining the electric pulse

that will be delivered to specific well sets or wells on an electroporation plate

Waveform: defines the type of electric pulse delivered to the cells

n

The exponential waveform builds up a charge in a capacitor and when the charge
is applied to the sample the voltage delivered decays exponentially, until the charge
remaining is about 37% of the original pulse

n

The square waveform relies on a charge being applied to the cells for a set time

Well set: a set of four adjacent wells in a column on a 96-well plate; the same

electroporation conditions or protocol are applied to all the cells of a well set

Whole plate protocol: a preprogrammed set of 24 protocols delivered across

the entire plate

Abbreviations

Cgrad: capacitance gradient

D: duration of pulse

Dgrad: duration gradient

Exp: exponential

NP: number of pulses

P: pulse

Sqr: square

Vgrad: voltage gradient

Visit us on the Web at discover.bio-rad.com | Electroporation Optimization Glossary

3

Gene Pulser MXcell Electroporation Guide

Electroporation Parameters Selection Pathways

The first parameter to identify is the waveform. This guide will take you down two
possible paths to identify other parameters that will yield the best transfection results.
These parameters can be identified in parallel using the MXcell system.

Identify Pulse Waveform

Exponential Waveform Square Waveform

Optimize Capacitance Optimize Voltage

Optimize Voltage Optimize Pulse Duration

Optimize Resistance Optimize Number of Pulses

Path taken to determine optimal electroporation conditions

4

Visit us on the Web at discover.bio-rad.com | Electroporation Optimization Pathways

Gene Pulser MXcell Electroporation Guide

Start Your Protocols Selection Here

To choose a starting

set of protocols,

search our library of

electroprotocols at

www.bio-rad.com/

genetransferprotocols/

or search

http://www.ncbi.nlm.

nih.gov/PubMed/.

Known electroporation conditions

You know the electroporation conditions or found
a suitable set of protocols in our library

You need a quick confirmation that your existing
protocol is optimal

You know the waveform

Unknown electroporation conditions

You are working with a new cell line or cell type

You have no information about electroporation conditions

Go to page 6

Go to page 7

Visit us on the Web at discover.bio-rad.com | Protocols Selection

5

Gene Pulser MXcell Electroporation Guide

Known Electroporation Conditions

Cell number

Based on the

number of cells

availabl e for your

experime nt, you

will use eithe r a

whole or a partia l

electro poratio n

plate (4– 6 well sets).

Cell number <5 x 10

6

Use Mini Protocols

Cell number >5 x 10

6

Use Whole Plate Protocols

Multiple cell lines

Use Whole Plate Protocols

Go to pages

9–10

Go to pages

11–12

Go to page 13

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Visit us on the Web at discover.bio-rad.com | Protocols: Known Conditions