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BTX ECM 830, User Manual

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Publication 5423-020-REV 1.0   •   www.btxonline.com

Table of Contents

SUBJECT

PAGE #

General Optimization Guide for Electroporation

36 – 38

  DNA Delivery into Cells Using Electroporation

36

  Wave Forms

37

  Field Strength

37

  Pulse Length

38

  Number of Pulses

38

  Electroporation Buffer

38

  DNA/RNA Concentrations

38

Applications

39 – 41

  Bacteria and Yeast Transformation

39

  Plant and Insect Transfection

39

Electroporation Buffers

40

Ordering Information

41

Summary of Contents for ECM 830

Page 1: ...Publication 5423 020 REV 1 0 Exponential Decay Wave Electroporation System USER S MANUAL ...

Page 2: ...ptly contact BTX 84 October Hill Road Building 7 Holliston Massachusetts USA 01746 1388 using our toll free number 1 800 272 2775 US Only or 508 893 8999 E mail support hbiosci com Goods will not be accepted for return unless an RMA Returned Materials Authorization number has been issued by our customer service department The customer is responsible for shipping charges Please allow a reasonable p...

Page 3: ...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 specifications if used in applications on...

Page 4: ... team constantly tracks published literature for any reference to electroporation and electrofusion We extract the pertinent experimental conditions and yields from these papers to help us in our efforts to help you In addition to tracking publications we are available to you for support at any time for advice in experimental design product recommendations troubleshooting and any other relevant te...

Page 5: ...g Experiments 15 17 Creating New Protocols 21 Using Specialty Protocols 18 20 Using an HT Plate Handler 18 19 Using Specialty Electrodes 20 Managing Protocols 21 23 Saving a Copy of a Protocol 21 Renaming a Protocol 21 Deleting a Protocol 22 Password Protecting a Protocol 23 Protocol Manager Software 24 26 Installation 24 Overview 24 Upload from Generator to PC 24 Download from PC to Generator 25 ...

Page 6: ...ration 36 38 DNA Delivery into Cells Using Electroporation 36 Wave Forms 37 Field Strength 37 Pulse Length 38 Number of Pulses 38 Electroporation Buffer 38 DNA RNA Concentrations 38 Applications 39 41 Bacteria and Yeast Transformation 39 Plant and Insect Transfection 39 Electroporation Buffers 40 Ordering Information 41 ...

Page 7: ...n electroporation applications The electrical pulse data is captured as both a graphic display of the waveform and electrical output values following each experiment The ECM 630 is an exponential decay wave electroporation generator providing a broad range of voltage and time constants for full flexibility in varying transfection applications The ability to select the resistance and capacitance va...

Page 8: ...on Once you have determined that the components of the system have not sustained any obvious damage in shipment proceed with the installation Remove the insulating tab from the battery at the base of the unit Place the generator in a location 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 U...

Page 9: ...ple prior to delivering the DC pulse Run Protocol Used to deliver the pulse protocol to the sample Settings Also referred to as Gear icon Access the settings menu used to adjust the following parameters Date and Time Audible Alarms Backlighting and software updates Displays device information Stop Protocol Used during the pulse delivery sequence to stop the progress of the protocol Protocol Locked...

Page 10: ...keypad to type the time Tap the Green Check icon to save and return to the settings screen Setting Audible Alarm Preferences On the home screen tap the Gear icon Tap Audible Alarms icon Select preferred audible alarms by tapping the icons to activate or deactivate Tap the Green Check icon to save and return to the settings screen Software Setup NOTE You may change the date and time format by tappi...

Page 11: ...Tap Device Information icon Select preferred brightness by tapping the icon indicating brightness Tap the Green Check icon to save and return to the settings screen View device information Tap the Green Check icon to return to the settings screen 1 1 6 8 7 Displaying Device Information On the home screen tap the Gear icon Software Setup ...

Page 12: ...ired cells 13 74 73 Using Preset Protocols continued 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 for future use You may continue pulsing go back to the protocol select screen or to the home screen Preset Protocols...

Page 13: ...the Preset Protocols icon Double tap desired cell type Scroll through available cells Tap desired cells Tap Folder icon Customizing a Preset Protocol continued 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 16 17 Preset Protocols ...

Page 14: ...t Protocol continued Scroll through user protocols to locate the newly saved method Double tap the newly saved method Tap the parameter s requiring customization Proceed to modify selected parameters Tap the Green Check icon to proceed to the run screen Preset Protocols ...

Page 15: ...e Green Check icon to save Creating New Protocols On the home screen tap User Protocols icon 17 17 24 Creating New Protocols On the home screen tap User Protocols icon 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 ...

Page 16: ... values Tap the Number of Pulses Pulse Interval icon Use the numeric keypad to set the number of pulses Creating New Protocols continued Tap the Resistance Capacitance icon 26 29 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 Intervals must only be set when performing multiple pulsing ...

Page 17: ...mega icon 32 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 for future use You may continue pulsing go back to set up screen or to the home screen Performing Experiments NOTE See the Protocol Manager Software section for instruction...

Page 18: ...ge Tap the Green Go icon to run protocol The Red x icon can be used to stop the protocol at any time Using an HT Plate Handler Plug the HT Plate Handler into the front ports of the ECM 630 unit Select desired protocol preset protocol or user protocol Double tap desired cells 49 Using Specialty Protocols NOTE Generator will display readings after first and last pulse of each column NOTE Note the me...

Page 19: ...being electroporated To run tap the Omega icon 57 80 81 55 56 With load measurement OK press the Green Go icon to run protocol While the pulse is being delivered the Red x icon can be used to stop the protocol at any time 79 Using an HT Plate Handler continued Once protocol is complete data regarding your pulse is displayed and stored in the systems logs for future use You may continue pulsing go ...

Page 20: ...ecialty Electrodes Once protocol is complete data regarding your pulse is displayed and stored in the systems logs for future use You may continue pulsing go back to the settings screen or the protocol select screen or to the home screen Using Specialty Protocols ...

Page 21: ...ame for your protocol using the onscreen keyboard Choose the Symbols Numbers icon 123 to display the list of available numbers and symbols that can be included in the protocol name To switch the keyboard back to letters mode choose the Letters icon ABC 15 characters maximum 4 When finished entering the new Protocol Name choose the Green Check icon to save your changes 58 Renaming a Protocol Follow...

Page 22: ...en tap the User Protocols icon Tap the protocol you wish to delete Tap Folder icon Tap Delete icon Confirm delete by tapping the Delete icon again Deleting a Protocol 17 Protocol has been removed from the menu Managing Protocols NOTE Preset protocols cannot be deleted ...

Page 23: ... wish to password protect View the protocol Tap the Open Lock icon 17 Confirm password protection by tapping the Open Lock icon again Use the numeric keypad to set the password Tap the Green Check icon to save The protocol is now locked against modifications To unlock press the Lock icon Type in the password then Tap the Green Check icon Managing Protocols ...

Page 24: ...rview Protocol Manager allows the user to Upload protocols from the ECM 630 generator to a PC Download protocols from a PC to the ECM 630 generator Upload log files from the ECM 630 generator to a PC Log File Control PC to Generator Protocol Control Generator to PC Protocol Control Upload from Generator to PC 1 Select Protocol s 2 Click Save Protocol 3 Select the drive and folder location 4 Click ...

Page 25: ...tor 1 Click Choose Protocol 2 Select protocol s to download 3 Click Open 4 Protocol name being downloaded is shown Protocol Manager Software NOTE If a protocol already exists on the ECM 630 generator Protocol Manager will not overwrite the file if you attempt to re download it ...

Page 26: ...le generator is running Program works best while generator is idle on protocol setup screen 2 Order of Operations A Connect USB B Turn generator on C Start Protocol Manager program Protocol Manager Software NOTE Log Files are saved as a tab delimited text file They can be opened with any common text editor like Notepad or by using spreadsheet software such as Excel NOTE Files should be backed up p...

Page 27: ...B open the Control Panel and select System and Security and then System Click Device Manager on the left hand menu Under Ports search for the USB serial port for ECM 630 This will show the COM Port to which the generator is connected as shown below Once the COM port is known open the Remote Screen Interface application Choose the correct COM port and click Connect You should now be able to control...

Page 28: ...oltage Width Droop and Sample Resistance Display 7 0 in color display Controls Touch Screen Programmability Storage over 1000 Protocols Arc Control Yes Other Specifications Interlock Switches Yes Pre Pulse Sample resistance check Yes Pulse Over Current Protection Yes PC control Yes PC communications Yes Log Report Yes Remote Operation Yes Foot Switch Control Yes Power Ratings 35 W idle and 350 W p...

Page 29: ...e servicing the fuse 2 Use only Type 3AG 1 4 x 11 4 in 5 amp 250 volt Type T time delay fuses Turn off power and remove power cord from power module Use a straight blade screwdriver to pry open the access door Remove the fuse holder and then remove the fuses from this holder as shown in figure below Replace fuses and then replace the fuse holder Battery Replacement Underside of Instrument 1 Discon...

Page 30: ...lied with the ECM 630 1 Connect the ECM 630 generator to the computer via USB Open the control panel and select System and Security and then System Click Device Manager on the left hand menu Pin Signal 1 5V 2 Data 3 Data 4 GND Mating Face 2 Two Flash update applications will be displayed in the device manager under Other Devices Right click on either of the flash update applications and select Upd...

Page 31: ...rding the driver publisher is displayed choose Install this driver software anyway USB Virtual CommPort Driver Installation continued 4 Download the software from BTX website www btxonline com Click on the Technical Resources drop down menu and select Downloads Download and install file named Firmware Updater Upgrading ECM 630 Series Software ...

Page 32: ...cal Resources drop down menu and select Downloads Download and install file named Firmware Updater 5 Click Open Update File and browse to the BTX Gemini 7 vx x x srec file that was previously saved to the desktop 6 Click Start Update 7 After the update is complete press Reboot and then power cycle the ECM 630 generator from the power switch in the back of the unit 8 Repeat steps 5 through 7 with t...

Page 33: ...ge selected 2 Gap size is too small for the voltage selected 3 Pulse duration is too long No_HB_from_GUI Communication error Hardware error Please contact the manufacturer Sample Resistance Out Of Range The measured sample load is out of range Increase the sample resistance This can be achieved by 1 Increasing the gap size 2 Reducing the volume 3 Replacing the buffer with less conductive type of b...

Page 34: ...stored an on screen message is displayed to indicate the interruption Out of Range A value was entered or encountered in a protocol that was beyond the generator s limits 71 Low Battery Indication Time Date area will flash between Date and Time and LOW BATTERY message will display 72 83 Troubleshooting NOTE It is recommended that you save your protocols periodically by uploading them to a PC ...

Page 35: ...s Is the voltage correct Chamber gap Pulse length or appropriate instrument settings Number of pulses If so follow Optimization Guidelines outlined Low Viability Verify physical biological chemical parameters Are the voltage chamber gap pulse length time constant pulse number and other instrument settings correct If so reduce voltage pulse length or number of pulses and reoptimize protocol to impr...

Page 36: ...in experimental conditions and parameters of electrical pulses may be capable of causing many more molecules to move per unit time than simple diffusion There is also good evidence Sukharev et al 1992 that DNA movement is in the opposite direction An additional important consideration when the voltage pulse is applied to the cells and medium is that the amount of current that flows is dependent on...

Page 37: ...red for optimizing field strength 1 Electrode Gap Size 2 Cell Diameter 3 Temperature Cell Type Field Strength Ranges Bacteria Yeast 3 24 kV cm Mammalian 0 25 3 kV cm Plant 3 12 kV cm Electrode Gap Size The distance between electrodes or gap size is important when optimizing your electroporation experiment Field strength is calculated using voltage divided by gap size For example using a 4 mm gap c...

Page 38: ... water shortly before pulse This swelling of the cells results in lowering the optimal permeation voltage while ensuring the membrane is more easily permeable for many cells but can be damaging to others Prokaryotic cells such as bacteria require the use of high resistance buffers 3000 ohms For this reason proper preparation and washing of the cells is essential to remove excess salt ions to reduc...

Page 39: ...ts µg DNA are commonly seen for gram negative bacteria while for gram positive bacteria generally 1x106 transformants µg DNA are achievable Plant and Insect Transfection Electroporation of plant tissue can be used to generate transgenic crops that are useful in agricultural horticultural applications Insect models are also widely used throughout the scientific community to study development and ge...

Page 40: ...compared to bicarbonate buffers which are also commonly used in cell culture RPMI a form of medium used in cell culture and tissue culture It has traditionally been used for growth of Human lymphoid cells This medium contains a great deal of phosphate and is formulated for use in a 5 carbon dioxide atmosphere Opti MEM an improved Minimal Essential Medium MEM that allows for a reduction of fetal bo...

Page 41: ...tions 1 bottle 45 0805 BTXpress Solution 10 0 ml Bottle For up to 100 Reactions 1 bottle Order No Description Qty Accessories 45 0110 Flatpack Chambers 0 56 mm gap 80 μl volume pkg of 50 Requires Safety Stand 45 0207 or 45 0208 Pkg of 50 45 0109 Flatpack Chambers 1 83 mm gap 1 5 ml volume pkg of 50 Requires Safety Stand 45 0207 or 45 0208 Pkg of 50 47 0206 Flatpack Chambers 4 mm gap 10 ml volume p...

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