Mirus Ingenio EZporator MTR 51000 Operational manual

Ingenio® EZporator®
Electroporaon System
Product User Manual
MIR 51000
RevB 121520
mirusbio.com

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Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Table of Contents
Technology Overview..................................................................... 3
System Components and Features.................................................. 4
Technical Specicaons.................................................................. 5
Safety Informaon—Please Read Carefully..................................... 6-7
Set-up Instrucons
Installaon.................................................................................. 8
Inializing.................................................................................... 8
Instrument Features and Controls................................................ 8
Display and Status Opons.......................................................... 9
Post-pulse Display....................................................................... 10
Voltage Adjust Knob, Pulse Buon………....................................... 10
EZporator® System Protocols
Quick Reference Guidelines......................................................... 11
Instrucons for Use with Ingenio® Soluon and Kits..................... 12-13
Opmizaon
Cell Density and Passage Number................................................ 14
Nucleic Acid Purity and Concentraon......................................... 14
Pulse Condions.......................................................................... 15
Pulse Condions for Common Cell Types …………….......................... 16
Instrument Troubleshoong........................................................... 17
Experimental Troubleshoong
Low Electroporaon Eciency..................................................... 18
High Cellular Toxicity................................................................... 19
Maintenance.................................................................................. 20
Warranty
General Warranty........................................................................ 21
Out of Warranty Service.............................................................. 22
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Mirus Bio LLC
www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852
Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Technology Overview
Electroporaon is a physical transfecon method that ulizes short electrical pulses
to create transient membrane pores in cells through which small parcles (e.g. DNA,
RNA, proteins, etc.) can pass. Opmal electroporator pulse sengs and composion
of cell resuspension soluon greatly enhance electro-permeabilizaon and promote
passage of parcles through the cell membrane. Since electroporaon is an eecve
transfecon method regardless of the type of molecule delivered or the target cells,
this technique provides a robust and universal approach for mulple applicaons,
including gene expression, RNAi, stable cell line generaon and CRISPR/Cas genome
eding.
Electroporaon is an invaluable alternave for cell types that are not responsive to
chemical transfecon and oen yields signicantly higher transfecon eciency.
Opmizing parameters such as nucleic acid amount, cell density and voltage for
each cell type will improve transfecon eciency and decrease cell mortality, thus
making electroporaon suitable for transfecng various cell types with any type of
nucleic acid or small molecule.
The Ingenio® EZporator® Electroporaon System was developed with both eciency
and simplicity for the researcher in mind. This system ulizes an exponenal decay
pulse* to eecvely deliver cargo to cells in a cuvee with the push of a buon. The
Ingenio® EZporator® was designed for use with Ingenio® Electroporaon Kits and
Soluon. Use of electroporaon soluons not manufactured by Mirus Bio may result
in low transfecon eciency and high cellular cytotoxicity. Addionally, use of in-
compable electroporaon soluons may cause damage to the instrument and will
void the warranty. More informaon on Ingenio® Electroporaon Soluon and Kits
is available at mirusbio.com/ingenio.
*In exponenal-decay pulse condions, a set voltage is released from the capacitor and decays
exponenally. See Opmizaon Secon (page 14) for addional details.
Ingenio® EZporator® Electroporaon System and Ingenio®
Electroporaon Soluon, cuvees and cell droppers.

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System Components and Features
The EZporator® Electroporaon System was designed for mammalian and insect cell
transfecon with Ingenio® Electroporaon Kits and Soluon.
System features include:
• Digital Liquid Crystal Display (LCD) user interface
• Pulse Generator to deliver exponenal decay pulse at user-determined voltage.
NOTE: In exponenal decay pulse condions, a set voltage is released from the
capacitor and decays exponenally.
• Cuvee Chamber compable with 0.1 cm, 0.2 cm or 0.4 cm cuvees
• Power cords with Type B, F and G electrical outlet plugs
• Low Voltage (LV) mode
- Voltage range of 20 to 400 V with 2 V resoluon
- 150 Ω internal resistance and 1,050 µF capacitance
• High Voltage (HV) mode
- Voltage range of 30 to 2,500 V with 10 V resoluon
- 150 Ω internal resistance and 36 µF capacitance
• Streamlined experimental approach: (1) Inialize instrument, (2) Set voltage
and mode with the Voltage Adjust knob and (3) Deliver electroporaon pulse
by pressing the Pulse buon.
• Dual monitoring and display of peak output voltage and me constant
System Components:
Upon receipt, carefully open the box containing the EZporator® Electroporaon
System and verify that the box contains the following:
• EZporator® Pulse Generator (1)
• EZporator® Cuvee Chamber (1)
• Power cords (3)
Mirus Bio LLC
www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852
Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
EZporator® Pulse Generator Power Cords
EZporator® Cuvee Chamber

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Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Display Type: 16-character LCD backlit
Dimensions (W×D×H) 254 mm × 254 mm × 140 mm (Pulse Generator)
Power Source
Voltage 100 to 240 VAC, 50-60 Hz, CAT I
Power 800 W
Fusing 2.5 A, T rang 250 V
Environmental Usage:
Operang Temperature 10°C to 40°C (NOTE: Intended for indoor use only)
Cooling Convecon through metal case
Relave Humidity Avoid excessively humid working environments
Altude < 2,000 m above sea level (operang)
Mechanical Characteriscs:
Maximum Voltage Output: 2,500 V peak
Maximum Pulse Length 125 ms at 400 V peak or 5 ms at 2,500 V peak
Pulse Waveform Exponenal decay
Resistance 150 Ω internal resistor
Polluon Degree 2 Not to be operated in conducve pollutants atmosphere
Ordering Informaon:
Catalog # Product
MIR 51000 Ingenio® EZporator® Electroporaon System
MIR 51100 Ingenio® EZporator® Pulse Generator
MIR 51200 Ingenio® EZporator® Cuvee Chamber
Mirus Bio LLC
www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852
Technical Specicaons

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www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852
Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
General Safety Summary
The EZporator® Electroporaon System is safe to use when operated in accordance
with this manual. Please read the following safety precauons to ensure proper use,
as this system is designed to deliver high voltage electrical pulses. If the equipment
is improperly used, the protecon provided by the equipment may be impaired.
To prevent hazard or injury, take the following precauons:
• USE PROPER POWER CORD: Use only the power cord specied for this product
and cered for the country of use.
• USE PROPER FUSE: Use only 5 mm × 20 mm, 2.5 A, 250 V type T (me delay) fuses
with this product.
• ORIENT THE EQUIPMENT PROPERLY: Do not orient the equipment such that it
is dicult to manage the connecon and disconnecon of devices.
• ENSURE PROPER VENTILATION: Allow 3 to 5 cm of clearance on all sides of the
module for proper cooling.
• GROUND THE PRODUCT: This product is grounded through the grounding con-
ductor of the power cord. To avoid electric shock, the grounding conductor
must be connected to earth ground. Before making any connecons to the
input or output terminals, ensure that the EZporator® is properly grounded.
• OBSERVE ALL TERMINAL RATINGS: Review the user manual to learn the rangs
on all connecons.
• AVOID CONTACT WITH INTERNAL CIRCUITRY: Do not open the product or
touch any electronic circuitry inside of the product.
• AVOID EXPOSURE TO EXCESS MOISTURE: Do not expose the device to a hu-
midied environment. If uid is spilled on or near the EZporator® instrument,
performance and safety may be compromised.
• DO NOT OPERATE IF DAMAGED: If damage is suspected on or to the EZpora-
tor® Pulse Generator or Cuvee Chamber, contact qualied service personnel
to perform an inspecon prior to use.
• OBSERVE ALL WARNING LABELS ON PRODUCT: Read all labels on product to
ensure proper usage.
• ENVIRONMENTAL USAGE: This product should not be used in the presence of a
ammable atmosphere such as an anesthec mixture with air, oxygen or ni-
trous oxide.

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Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
General Safety Summary (Connued)
High Voltage Risk
This symbol indicates a risk of electric shock. Electric shocks are dangerous
and may cause personal injury or death.
This instrument contains a high voltage power supply adjustable to 2,500 V. This
presents a serious risk of personal injury if not used in accordance with design and/
or use specicaons, if used in applicaons or with products for which they are not
intended or designed, or if they are used by untrained or unqualied personnel.
Please take special note of the following:
• The user must read this manual carefully before operang the instrument.
• Removing the EZporator® Pulse Generator cover and breaking the “Warranty
Void if Seal Broken” scker will void the warranty.
• Power o the instrument before connecng or disconnecng any cords/cables.
• Do not open the Cuvee Chamber or aempt to touch the electrodes while the
electrical pulse is being applied.
• All service must be performed by Mirus Bio authorized personnel only.
If there are any quesons about the operaon of this instrument, call Mirus Bio
Technical Support at 888.530.0801 or +1.608.441.2852.
Cauon Noce
This symbol indicates that cauon is advised. Proper set-up, usage and PPE
when using this instrument are required.
The Ingenio® EZporator® Electroporaon System is intended for laboratory use only
and can be used in research and development applicaons. These systems have
been designed to meet the standards for electromagnec compability (EMC) in-
tended for laboratory equipment applicaons as well as the applicable safety re-
quirements for electrical equipment for measurement, control and laboratory use.
The unit itself does not generate waste, but may be used to treat samples that are
hazardous. Please use appropriate PPE and ensure disposal in accordance with local
regulaons and pracces.
This product should NOT be used in the presence of a ammable atmosphere such
as an anesthec mixture with air, oxygen or nitrous oxide.
This product is for RESEARCH USE ONLY and is not intended for clinical use on
animals or human paents.
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www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852

Page 8
Mirus Bio LLC
www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852
Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Set-up Instrucons
Installaon
Place the EZporator® Pulse Generator on a laboratory bench or work table. Be sure
to allow 3 to 5 cm of clearance on all sides of the module for proper cooling. When
selecng a workspace, choose a stable, dry locaon with an easily accessible power
outlet. NOTE: It is normal for the EZporator® Pulse Generator module to be slightly
warmer than its operang environment.
Connect the power cord to the EZporator® Pulse Generator module AC Input on the
back panel, then insert the power cord plug into an appropriate electrical outlet (i.e.
110 - 240 VAC).
To connect the EZporator® Cuvee Chamber to the Pulse Generator, insert the color
coded plugs aached to the Cuvee Chamber into the HV Output located on front
panel of the EZporator® Pulse Generator.
Inializing
1. Turn the EZporator® Pulse Generator on by pushing the green power buon
located at the back of the instrument (see page 19). NOTE: Push the power
buon once to turn on. Push again to turn o. Do not hold the power buon.
2. The EZporator® will go through a series of self-test algorithms to test generator
funconality. The display will ash “EZporator®” and the soware version (e.g.
V1.01) prior to aaining a ready status.
3. When the instrument is inialized for the rst me, the display will read
“READY LV 400V.”
Instrument Features and Controls:
Cuvee Chamber High Voltage
Output
LCD
Pulse Buon
Voltage Adjust

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Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Set-up Instrucons (Connued)
Display
Once inialized, the EZporator® Pulse Generator display will show the operaonal
status, voltage mode (i.e. LV or HV) and set voltage. Once a pulse is delivered to the
cuvee containing cells, the display will read the peak output voltage delivered and
the me constant.
Status opons include:
Ready Mode
The EZporator® is operaonal and ready to deliver a pulse. The
LCD display will show HV for the high voltage mode and LV for
low voltage mode. NOTE: Ensure that the correct voltage mode
is selected for your cell type and electroporaon buer. Use LV
mode for mammalian cell types in Ingenio® Soluon.
• LV mode: from 20 to 400 V (2 V increments), 1050 µF capacitance
• HV mode: from 20 to 2500 V (10 V increments), 6 µF capacitance
Inializing and Charging
Inializing and charging capacitor to the preset voltage level.
Pulsing
Delivering a voltage pulse.
Feedback
Displays peak voltage and me constant for delivered pulse.
Sequence Aborted
Aborted delivery of a pulse at the user’s request. To abort a
pulse sequence, press the Pulse Buon a second me before
delivery of the pulse.
SCR Failure
The instrument has not detected an output pulse during the
pulsing operaon. As a safety precauon, this message can only
be reset by turning the power o.
Charge Failure
The capacitors failed to charge properly. To reset this message,
press the ‘Pulse’ buon.
Setpoint Too Low
The entered voltage setpoint is below the normal operang
specicaons. No pulse has been applied.
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Page 10
Ingenio® EZporator® Electroporaon System
Product User Manual | Instrucons for MIR 51000
Set-up Instrucons (Connued)
Post-pulse Display
The EZporator® instrument will display the peak discharge voltage (VP) in volts (V)
and the me constant (t) in milliseconds (ms). It is normal for the measured voltage
to dier slightly from the setpoint. This is due to internal resistance and will uctu-
ate with electrical load variaons (i.e. buer conducvity). Following the delivery of
a pulse, press the Pulse Buon or rotate the Voltage Adjust knob to return to the
ready mode.
NOTE: With exponenal decay generators such as the EZporator®, it is necessary to
idenfy and monitor the me constant (t), as this variable is dependent on the con-
ducvity and osmolarity of the electroporated sample. Signicant changes in the
electroporaon sample (e.g. variaons in cell density, volume of nucleic acid added,
electroporaon soluon used) will alter the me constant.
Voltage Adjust Knob
The EZporator® Voltage Adjust knob controls set voltage AND voltage mode (i.e. LV
and HV modes). There is no separate voltage mode switch. Use the Voltage Adjust
knob to adjust the voltage from 20 to 400 V in LV mode (in 2 V increments) and from
30 to 2,500 V in HV mode (in 10 V increments). If in LV mode, increasing of the volt-
age setpoint past 400 V will result in a switch to HV mode. The voltage then read-
justs to 0 V and connues up to 2,500 V. If in HV mode, decreasing the voltage set-
point past 0 V will result in a switch to LV mode. The voltage then readjusts to 400 V.
NOTE 1: The EZporator® emits a sound when switching between LV and HV modes.
NOTE 2: The Voltage Adjust knob is speed sensive. A quick rotaon increases the
rate of change of the voltage setpoint.
NOTE 3: LV mode is used for mammalian and insect cell types.
Pulse Buon
The Pulse buon is acvated in ‘READY’ mode. Once pressed, the generator will
bleed o the capacitors to the preset voltage, prior to delivering the pulse. The
capacitor bleed me is typically between 1-6 seconds. A pulse sequence may be
aborted by pressing the Pulse buon a second me before delivery of the pulse.
Following the delivery of a pulse, press the Pulse buon or rotate the Voltage Adjust
knob to return to the ‘READY’ mode.
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www.mirusbio.com │ [email protected] │ U.S. Toll Free: 844.MIRUSBIO (844.647.8724)│ Direct: +1.608.441.2852