BTX Gemini X2 User manual

5423-018 REV 1.0
Twin Waveform
Electroporation Systems
USER’S MANUAL

2Publicaon 5423-018 REV 1.0 • www.btxonline.com
Research Only
BTX
84 October Hill Rd
Holliston, MA 01746, USA
Phone: [phone]
Fax: [phone]
Web: www.btxonline.com
Warranty
BTX warranes the Gemini X2 and BTX Gemini SC Twin Waveform
Electroporaon Systems for a period of two years from the date
of purchase. At its opon, BTX will repair or replace the unit if
it is found to be defecve as to workmanship or materials. This
warranty does not extend to any instrumentaon which has been
(a) subjected to misuse, neglect, accident or abuse, (b) repaired
or altered by anyone other than BTX without BTX express and
prior approval, (c) used in violaon of instrucons furnished
by BTX. This warranty extends only to the original customer
purchaser. IN NO EVENT SHALL BTX BE LIABLE FOR INCIDENTAL OR
CONSEQUENTIAL DAMAGES. Some states do not allow exclusion
or limitaon of incidental or consequenal damages so the above
limitaon or exclusion may not apply to you. THERE ARE NO
IMPLIED WARRANTIES OF MERCHANTABILITY, OR FITNESS FOR
A PARTICULAR USE, OR OF ANY OTHER NATURE. Some states do
not allow this limitaon on an implied warranty, so the above
limitaon may not apply to you. Without liming the generality
of the foregoing, BTX shall not be liable for any claims of any kind
whatsoever, as to the equipment delivered or for non-delivery of
equipment, and whether or not based on negligence. Warranty
is void if the BTX Gemini X2 and BTX Gemini SC instrument is
changed in any way from its original factory design or if repairs
are aempted without wrien authorizaon by BTX. Warranty is
void if parts, connecons or electrodes not manufactured by BTX
are used with the BTX Gemini X2 and BTX Gemini SC instrument.
If a defect arises within the warranty period, promptly contact
BTX, 84 October Hill Road, Building 7, Holliston, Massachuses,
USA 01746-1388 using our toll free number 1-800-272-2775 (US
Only) or 508-893-8999 (E-mail: [email protected]). Goods will
not be accepted for return unless an RMA (Returned Materials
Authorizaon) number has been issued by our customer service
department. The customer is responsible for shipping charges.
Please allow a reasonable period of me for compleon of repairs,
replacement and return. If the unit is replaced, the replacement
unit is covered only for the remainder of the original warranty
period dang from the purchase of the original device. This
warranty gives you specic rights, and you may also have other
rights, which vary from state to state.
Out of Warranty Service
Proceed exactly as for Warranty Service above. If our service
department can assist you by phone or other correspondence, we
will be glad to help at no charge.
Repair service will be billed on the basis of labor and materials.
A complete statement of me spent and materials used will be
supplied. Shipment to BTX should be prepaid. Your bill will include
return shipment freight charges.
Disassembly by the user is prohibited. Service should only be
carried out by experienced BTX technicians.
Repair Facilies and Parts
BTX stocks replacement and repair parts. When ordering, please
describe parts as completely as possible, preferably using our part
numbers. If praccal, enclose a sample photo or drawing.
Cauon Noce
The BTX Gemini X2 and BTX Gemini SC systems are 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) and safety
intended for laboratory equipment applicaons.
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.
Warranty Information

3Publicaon 5423-018 REV 1.0 • www.btxonline.com
Please read the following safety precauons to ensure proper
use of your generator. If the equipment is used in a manner not
specied, the protecon provided by the equipment may be
impaired.
To Prevent Hazard or Injury
Use Proper Line Cord
Use only the specied line cord for this product and make sure
line cord is cered for country of use.The operang voltage range
for the BTX Gemini Twin Wave Series is 100–240 vac, 50/60 Hz.
Ground the Product
This product is grounded through the grounding conductor 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 of the product, ensure that the
product is properly grounded.
Make Proper Connecons
Make sure all connecons are made properly and securely.
Any signal wire connecons to the unit must be no longer than
three meters.
Observe All Terminal Rangs
Review the operang manual to learn the rangs on all
connecons.
Use Proper Fuse
Use only specied fuses with product.
Avoid Exposed Circuitry
Do not touch any electronic circuitry inside of the product.
Do Not Operate with Suspected Failures
If damage is suspected on or to the product do not operate
the product. Contact qualied service personnel to perform
inspecon.
Orient the Equipment Properly
Do not orient the equipment so that it is dicult to operate the
disconnecon device.
Place Product in Proper Environment
Review the operang manual for guidelines for proper operang
environments.
Observe All Warning Labels on Product
Read all labels on product to ensure proper usage.
Caution Risk of
Electric Shock
Caution Protective Ground
Terminal
High Voltage Risk
These instruments contain a high voltage power supply adjustable
to 3,000 V. High voltage power supplies present a serious risk of
personal injury if not used in accordance with design and/or use
specicaons, if used in applicaons on products for which they
are not intended or designed, or if they are used by untrained or
unqualied personnel.
• The user must read this manual carefully before the
instruments are placed into operaon.
• Removing the cover will void the warranty.
• Do not connect or disconnect the high voltage cable with
the high voltage enabled.
• To connect or disconnect the cable, turn line power o and
unplug line (mains) cord.
• Do not touch the electrode p while the waveforms are
being applied.
If there are any quesons about the operaon of this instrument,
call BTX Customer service at 1-800-272-2775, or 1-508-893-8999.
Cauon Noce
The BTX Gemini Series Twin Waveform Electroporaon Systems
are 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)
intended for laboratory equipment applicaons as well as the
applicable safety requirements 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.
Safety Information

4Publicaon 5423-018 REV 1.0 • www.btxonline.com
Thank you for invesng in a Gemini System.
Since its founding in 1983, the main focus of BTX has been in the area of applying
controlled electric elds for genec engineering applicaons. Because of this,
we quickly established a reputaon as the technological leader in the elds of
electroporaon and electrofusion. Our systems have been installed in many
presgious instutes around the globe where they are used successfully for high
eciency transfecon, transformaon and cell fusion applicaons. We oer a
variety of waveforms, electrodes and chamber opons to provide you with the
best tools to achieve your goals.
We are vested in your success. To that end, the BTX technical support team
constantly tracks published literature for any reference to electroporaon and
electrofusion. We extract the pernent experimental condions and yields
from these papers to help us in our eorts to help you. In addion to tracking
publicaons, we are available to you for support at any me for advice in
experimental design, product recommendaons, troubleshoong, and any other
relevant technical advice.
We thank you again for your investment and we look forward to assisng you in
any way we can.
Finally, please read this manual carefully before aempng to operate the
electroporaon system. If you have any quesons about the unit or about
parcular applicaons, please contact us:
BTX
84 October Hill Road
Holliston, MA 01746 USA
Toll Free: 1-800-272-2775
Internaonal Callers: 508-893-8999
Fax: [phone]
Web: www.btxonline.com
Email: [email protected]
For any customers outside the US or Canada, please call your local BTX dealer or
call us directly.
A Message from BTX

5Publicaon 5423-018 REV 1.0 • www.btxonline.com
SUBJECT PAGE #
Warranty Informaon 2
Safety Informaon 3
Introducon 4
Table of Contents 5–6
Product Overview 7
Unpacking the System 8
Touchscreen Buon Reference 9
Soware Setup 10–11
Seng Time/Date 10
Seng Audible Alarm Preferences 10
Seng Backlight Preferences 11
Displaying Device Informaon 11
Preset Protocols 12–14
Using Preset Protocols 12
Customizing a Preset Protocol 13–14
Performing Experiments 15–21
Creang New Protocols 15
Creang an Exponenal Decay Wave Protocol 15–17
Creang a Square Wave Protocol 18–21
Using Specialty Protocols 22–24
Using an HT Plate Handler 22–23
Using Specialty Electrodes 24
Managing Protocols 25–27
Saving a Copy of a Protocol 25
Renaming a Protocol 25
Deleng a Protocol 26
Password Protecng a Protocol 27
Protocol Manager Soware 28–30
Installaon 28
Overview 28
Upload– Generator to PC 28
Download– PC to Generator 29
Upload Log Files 30
Remote Control Soware 31
Generator Specicaons 32–33
Maintenance 34
Upgrading Gemini Series Soware 35–37
USB Virtual Commport Driver Installaon 35–36
Device Updater Program 37
Error Messages & Troubleshoong 38–40
Electroporaon Overview 41
Table of Contents

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Table of Contents
SUBJECT PAGE #
General Opmizaon Guide for Electroporaon 42
Wave Forms 42
Field Strength 42
Pulse Length 43
Number of Pulses 43
Electroporaon Buer 43
DNA/RNA Concentraons 43
Applicaons 44
Mammalian Cell Transfecon 44
In Vivo, In Utero, In Ovo 44
Bacteria and Yeast Transformaon 44
Plant and Insect Transfecon 44
Electroporaon Buers 45
Ordering Informaon 46–47

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The BTX Gemini SC is a twin waveform generator incorporang
both square and exponenal decay waves in a single unit.
These waveform combinaons enable researchers to easily
and eciently electroporate eukaryoc and prokaryoc cells in
suspension with one easy-to-use setup. The BTX Gemini SC boasts
a wide range of voltage (10 to 3000 V, 5 V or 10 V increments),
pulse length (50 µs to 5 ms), me constant opons (which include
capacitance choices from 25 to 3275 µF in LV and 10, 25 or 50
µF in HV), mulple pulsing opon with the square waveform and
unparalleled pulse delivery accuracy.
The BTX Gemini SC, with unlimited custom protocol storage,
monitors and displays pre-pulse sample resistance as well as
delivered voltage values. The generator is controlled through a
color LCD touch screen interface. The enhanced safety features
of the BTX Gemini SC protect users as well as precious samples.
The Safety Dome allows researchers to safely work with
electroporaon cuvees, up to two at a me, giving researchers
the ability to experiment on sample volumes from 20 µl up to
800 µl.
This aordable system comes complete with the dual waveform
generator for suspension cell electroporaon, dozens of
preprogrammed protocols for commonly electroporated cells,
Safety Dome, 30 cuvees, cuvee stand, user manual, two-year
warranty, unlimited applicaon support and the same high quality
researchers have come to expect from BTX.
What is the dierence between
the Gemini X2 and the Gemini SC?
The Gemini X2 is designed to give researchers the ulmate control
and exibility in their experiments. It is possible to perform
electroporaon on ssues and organs in vivo, in utero, in ovo and
ex plant, on adherent cells, and on cells in suspension, in either
single cuvees or 96-well plates. It can be controlled remotely via
footswitch or PC, and it oers storage of pulse data. Because of
the Gemini X2 broad range of use, the specicaons are wide-
ranging, making the Gemini X2 the most versale electroporaon
system available today.
The Gemini SC is designed for researchers working to
electroporate cells in suspension in cuvees. This system cannot
accommodate mulple pulsing with the exponenal decay
waveform, remote operaon, specialty electrodes or 96-well
opons. For this reason, some of the specicaons of the Gemini
SC are not as extensive as what is available in the Gemini X2.
The BTX Gemini X2 is a highly advanced twin waveform generator
incorporang both square and exponenal decay waves in a single
unit. The BTX Gemini X2 has been designed with these waveform
combinaons to enable researchers to easily and eciently
electroporate eukaryoc cells and prokaryoc cells in all forms
with one easy-to-use setup.
The BTX Gemini X2, which can be operated via PC or remote
control, boasts a wide range of voltage (5 to 3000 V, 1 or 5 V
increments), pulse length (10 µs to 1s), me constant opons
(which include capacitance choices from 25 to 3275 µF in LV and
10, 25, 35, 50, 60, 75, 85 µF in HV), along with mulple pulsing
opons with both the square waveform and the exponenal decay
waveform, and unparalleled pulse delivery accuracy.
The BTX Gemini X2, with over 1,000 custom protocol storage,
monitors and displays pre-pulse sample resistance as well as
delivered voltage values and records logs of all experiment
parameters internally, which can be downloaded to a computer
for analysis and QC. The generator is controlled through a color
LCD touch screen interface and incorporates USB communicaons.
The pulse can also be acvated by a foot switch. The enhanced
safety features of the BTX Gemini X2 protect users as well
as precious samples. The BTX Gemini X2 is designed to give
researchers the ulmate exibility in their experiments, making it
possible to perform electroporaon on ssues and organs in vivo
(as well as in utero, in ovo, ex plant) on adherent cells, and cells in
suspension in either single cuvees or 96-well plates. The Safety
Dome allows researchers to safely work with electroporaon
cuvees, up to two at a me, giving researchers the ability to
experiment on sample volumes from 20 µl up to 800 µl.
This state-of-the-art system comes complete with the twin
waveform generator for cell electroporaon in all forms, dozens
of preprogrammed protocols for commonly electroporated cells,
Safety Dome, 30 cuvees, cuvee stand, user manual, two-year
warranty, unlimited applicaon support and the same high quality
researchers have come to expect from BTX.
Product Overview
CAUTION: FOR RESEARCH USE ONLY. NOT FOR
CLINICAL OR DIAGNOSTIC USE ON PATIENTS.

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The shipping carton in which your BTX Gemini Twin Waveform
Electroporaon System is packed has been specically designed
to provide maximum protecon to the instrument during
transportaon and normal handling condions. Upon receipt, the
carton should be examined for any external damage resulng from
shipment.
Open the carton and carefully remove the BTX Gemini Twin
Waveform Electroporator and inspect the unit for any apparent
damage. Save the carton and packing materials for future
transportaon and shipping requirements.
Packing Data
Check the packing slip to ensure that all items ordered and listed
are included in the shipment. Inform BTX immediately if any parts
are missing or damaged.
Power Source
As received, the instrument is ready for use with either 100–240 V
AC, 50/60 HZ.
The power requirements are 350 was. In the USA, the power
cord has a standard three prong plug.
Installaon
Once you have determined that the components of the system
have not sustained any obvious damage in shipment, proceed
with the installaon.
Remove the insulang tab from the baery at the base of the unit.
Place the generator in a locaon that is a dry, level, sturdy surface
free from extremes in ambient temperature, dust or chemical
exposures. Allow the device to equilibrate to room temperature
Unpack the safety dome, cuvee rack and disposable cuvee
chambers. Connect the safety dome, or in the case of the BTX
Gemini X2 system, specialty electrodes or HT plate handler, into
the connectors at the boom right-hand side of front panel.
Connect the mains/power cord to into the back panel at the
boom le.
Power up the system by pushing the rocker switch located on the
back panel at the boom le. The display will ash the BTX logo.
Once the soware inializes, the Main Menu screen will appear.
You are now ready to begin your work. Main Power Switch
Fuse Holder
Universal Power Input
USB Serial Input Footswitch Input
(switch sold
separately)
High Voltage Output
Remove Battery Insulating Tab.
Touchscreen Display
Unpacking the System

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Accept
Used to accept the parameters/sengs on a screen and
advance to the next screen in the menu, also used in
place of a double tap on various icons.
Back
Used to go back one screen.
Cancel
Used to cancel any changes/entries on a screen and
return to the previous screen in the menu.
Exponenal Decay Wave Indicator
Used to indicate when a protocol ulizes exponenal
decay wave pulses.
File Opons
Access the File Opon menu that is used to save,
rename and delete protocols.
Home/Main Menu
Access Preset Protocols, User Protocols and Sengs.
Page Down
Used to page down in a display list.
Page Up
Used to page up in a display list.
Scroll Down
Used to scroll down in a display list.
Scroll Up
Used to scroll up in a display list.
Pre-Pulse Resistance Measurement
Also referred to as Omega Icon. Used to measure the
resistance of the sample prior to delivering the DC
pulse.
Run Protocol
Used to deliver the pulse protocol to the sample.
Sengs
Also referred to as Gear icon. Access the sengs menu
used to adjust the following parameters: Date and Time,
Audible Alarms, Backlighng, and soware updates.
Displays device informaon.
Stop Protocol
Used during the pulse delivery sequence to stop the
progress of the protocol.
Square Wave Indicator
Used to indicate when a protocol ulizes square
wave pulses.
Protocol Locked
Indicates that changes to the protocol are currently
locked. Press the icon and enter password to unlock the
protocol.
Protocol Unlocked
Indicates that changes to the protocol are currently
unlocked. Pressing the icon twice will allow the user to
password protect the protocol.
Nocaon Box Quick Reference
Informaon / Status Message
Provides informaon or guidance for next steps,
including acceptable ranges for protocol programming.
Warning Message
Requires Nocaon Box tap for acknowledgement
of informaon, but does not prevent operaon of the
system.
Failure Message
Caused by user input error, failure of a protocol, or a
system error. Requires user to correct this issue in order
to complete the operaon.
Touchscreen Icons Reference

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Seng Time/Date
Aer inializing, the home screen will appear.
Tap the Gear icon.
Tap Time/Date icon.
Tap Date icon and use the keypad to type the date.
Tap Time icon and use the keypad to type the me.
Tap the Green Check icon to save and return to the sengs
screen.
Seng Audible Alarm Preferences
On the home screen, tap the Gear icon.
Tap Audible Alarms icon.
Select preferred audible alarms by tapping the icons to acvate
or deacvate.
Tap the Green Check icon to save and return to the sengs
screen.
Software Setup
NOTE: You may change the date and me format by tapping icons to
the right of the given values.

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Seng Backlight Preferences
On the home screen, tap the Gear icon.
Tap Backlight icon. Tap Device Informaon icon.
Select preferred brightness by tapping the icon indicang
% brightness.
Tap the Green Check icon to save and return to the sengs
screen.
View device informaon.
Tap the Green Check icon to return to the sengs screen.
Displaying Device Informaon
On the home screen, tap the Gear icon.
Software Setup

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Using Preset Protocols
On the home screen, tap the Preset Protocols icon.
Double tap desired cell type.
Scroll through available cells.
Tap desired cells.
Using Preset Protocols (connued)
Review parameters.
Tap Omega icon to measure pre-pulse load resistance.
With load measurement OK, press the Green ‘Go’ icon to run
protocol.
Once protocol is complete, data regarding your pulse is displayed
and stored in the systems logs (BTX Gemini X2 only) for future use.
You may connue pulsing, go back to the protocol select screen,
or to the home screen.
Preset Protocols

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Customizing a Preset Protocol
On the home screen, tap the Preset Protocols icon.
Double tap desired cell type.
Scroll through available cells.
Tap desired cells.
Tap Folder icon.
Customizing a Preset Protocol (connued)
Use keyboard to name copy of protocol.
Tap the Green Check icon to save.
Tap the Home icon.
On the home screen, tap User Protocols icon.
Preset Protocols

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Customizing a Preset Protocol (connued)
Scroll through user protocols to locate the newly saved method.
Double tap the newly saved method.
Tap the parameter(s) requiring customizaon.
Proceed to modify selected parameters.
Tap the Green Check icon to proceed to the run screen.
Preset Protocols

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Double tap New Protocol.
Use keyboard to name protocol.
Tap the Green Check icon to save.
Creang New Protocols
On the home screen, tap User Protocols icon.
Creang an Exponenal Decay Wave Protocol
Tap the Exponenal icon.
Tap the Green Check icon to save.
Double tap your newly saved protocol.
Tap the Voltage icon.
Use the numeric keypad to set voltage.
Tap the Green Check icon to save.
Performing Experiments

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Tap the Resistance (symbol) icon.
Use the numeric keypad to set resistance value.
Tap the Capacitance (µF) icon.
Tap the Number of Pulses / Pulse Interval icon.
Tap the Number of Pulses icon.
Use the numeric keypad to set the number of pulses.
Creang New Protocols (connued)
Creang an Exponenal Decay Wave Protocol
(connued)
Tap the Resistance / Capacitance icon.
Tap the Pulse Interval (sec) icon.
Use the numeric keypad to set the interval between pulses.
Tap the Green Check icon to save.
Performing Experiments
NOTE: LV and HV mode capacitance values dier. Use the numeric
keypad to set resistance value.
NOTE: The BTX Gemini X2 can perform up to 2 pulses with
Exponenal Decay Wave only if the Resistance > 100 ohms.
NOTE: Intervals must only be set when performing mulple pulsing.

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Use the numeric keypad to set the gap between electrodes.
Tap the Green Check icon to save.
Creang New Protocols (connued)
Creang an Exponenal Decay Wave Protocol
(connued)
Tap the Electrode Gap icon.
View your protocol.
To run, tap the Omega icon.
Tap the Green Check icon to proceed to the run screen.
With load measurement OK, press the Green ‘Go’ icon to run
protocol.
Once protocol is complete, data regarding your pulse is displayed
and stored in the systems logs (BTX Gemini X2 only) for future use.
You may connue pulsing, go back to set-up screen, or to the
home screen.
Performing Experiments
NOTE: See the Protocol Manager Soware secon for instrucons on
viewing and downloading log data.

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Creang New Protocols (connued)
Creang a Square Wave Protocol
On the home screen, tap User Protocols icon.
Performing Experiments
Double tap New Protocol.
Use keyboard to name protocol.
Tap the Green Check icon to save.

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Double tap your newly saved protocol.
Tap the Voltage icon.
Use the numeric keypad to set voltage.
Tap the Green Check icon to save.
Creang New Protocols (connued)
Creang a Square Wave Protocol
Tap the Square Wave icon.
Tap the Green Check icon to save.
Tap the Duraon icon.
Use the numeric keypad to set the pulse length.
Tap the Green Check icon to save.
Performing Experiments

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Tap the Number of Pulses icon.
Use the numeric keypad to set the number of pulses.
Creang New Protocols (connued)
Creang a Square Wave Protocol (connued)
Tap the Number of Pulses / Pulse Interval icon.
Tap the Pulse Interval (sec) icon.
Use the numeric keypad to set the pulse interval.
Tap the Green Check icon to save.
Tap the Electrode Gap icon.
Use the numeric keypad to set the gap between electrodes.
Tap the Green Check icon to save.
Tap the Green Check icon to proceed to the run screen.
Performing Experiments