Bio-rad Gene Pulser MXcell™ Electroporation System User Manual

Gene Pulser MXcell™ Electroporation SystemInstruction Manual Catalog #165-2670

Gene Pulser MXcell™ System Manual11 IntroductionCongratulations on the purchase of a Gene Pulser MXcell™ electroporation system! This instrument is

Gene Pulser MXcell™ System Manual | Introduction 2conditions can be programmed for use each time; the 12-well plate, 12 conditions. You can use the 96

Writing Conventions Used in This Manual3Writing Conventions Used in This ManualThis manual is for scientists and technicians who run the Gene Pulser

Gene Pulser MXcell™ System Manual | Introduction 4

Gene Pulser MXcell™ System Manual52 Get StartedThe Gene Pulser MXcell™ electroporation system ships as complete system that is ready to setup and st

Gene Pulser MXcell™ System Manual | Get Started6Setting Up the Gene Pulser MXcell SystemTo set up the Gene Pulser MXcell power module, follow these in

Setting Up the Gene Pulser MXcell System7The plate chamber holds the Gene Pulser electroporation plates. The top lid of the chamber must be closed t

Gene Pulser MXcell™ System Manual | Get Started8Introduction to the SystemYour Gene Pulser MXcell electroporation system is designed for ease of use a

Introduction to the System9Using the Control PanelThe control panel includes an LCD screen and a keypad: Figure 6. Gene Pulser MXcell system control

Gene Pulser MXcell™ System Manual | Get Started10Overview of Options in the Home ScreenThe Gene Pulser MXcell system home screen includes all the opti

Gene Pulser MXcellElectroporation SystemInstruction ManualBio-Rad Laboratories, Inc., 2000 Alfred Nobel Drive, Hercules, CA 94547 · 800-424-6723

Introduction to the System11To begin an operation on the Gene Pulser MXcell system, select one of the options in the home screen. Table 4 lists all

Gene Pulser MXcell™ System Manual | Get Started12Well Sets and Quadrants in Electroporation PlatesThe Gene Pulser electroporation plates are available

Well Sets and Quadrants in Electroporation Plates13When programming a protocol, the well set appears with the letters of the rows, followed by the c

Gene Pulser MXcell™ System Manual | Get Started14

Gene Pulser MXcell™ System Manual153 Prepare Mammalian CellsPreparation of cells is critical to the success of any electroporation experiment. This

Gene Pulser MXcell™ System Manual | Prepare Mammalian Cells16Harvesting and Counting the CellsTo harvest and count cells, follow these instructions:1.

Reagents and Solutions for Electroporation17Assessing Transfection EfficiencyTo assess the efficiency of a transfection experiment, several techniqu

Gene Pulser MXcell™ System Manual | Prepare Mammalian Cells18

Gene Pulser MXcell™ System Manual194 Program and Run the SystemThe Gene Pulser MXcell™ electroporation system runs experiments with Pre-Set protocol

Gene Pulser MXcell™ System Manual | Program and Run the System20Turning on the SystemTo turn on the Gene Pulser MXcell system, press the power button

iiCopyright ©2007 Bio-Rad Laboratories, Inc. Reproduction in any form, either print or electronic, is prohibited without written permission of Bio-Ra

Protocol Set-Up Option21Press the arrow keys to select a parameter, then press the alphanumeric keys to enter a new value. Press ENTER to confirm th

Gene Pulser MXcell™ System Manual | Program and Run the System225. Select a waveform (EXP or SQR). Press the arrow keys to select a waveform. Then en

Gradient Protocol Option23GRADIENT PROGRAMMINGTo program a gradient protocol, follow these instructions: 1. Select Gradient Protocol in the home scre

Gene Pulser MXcell™ System Manual | Program and Run the System245. (Optional) Select another quadrant.Press and release the up and down arrow keys to

Pre-Set Protocols Option254. Edit a selected protocol.Enter the changes and press the Save key to save the changes: • To rename a protocol, press t

Gene Pulser MXcell™ System Manual | Program and Run the System263. (Optional) Change the values of the parameters.Press the arrow keys to select the

Screen Intensity Option27• Press Delete to remove the selected protocol from the list.• At the screen prompt, press Yes to delete or No to return to

Gene Pulser MXcell™ System Manual | Program and Run the System28Saving ProtocolsOnce a protocol is programmed or a pre-set protocol is changed, you ha

Adding and Deleting the Users and Protocols296. Press the SAVE key to continue of the BACK key to the protocol.Tip: If a protocol is not saved and y

Gene Pulser MXcell™ System Manual | Program and Run the System30

iiiSafety and Regulatory ComplianceThe Gene Pulser MXcell™ electroporation system is designed to run safely. Please read the following sections to le

Gene Pulser MXcell™ System Manual315 Pre-Set ProtocolsThe Gene Pulser MXcell™ electroporation system provides a large set of pre-set protocols devel

Gene Pulser MXcell™ System Manual | Pre-Set Protocols32Refer to the page numbers listed in Table 6 (starting on page 32) for more information about ea

33Mixed protocols (page 38)13. Opt 96 well/ Exp,Sqr 96 Use with cell lines with no protocol reference. This protocol includes a range of average sta

Gene Pulser MXcell™ System Manual | Pre-Set Protocols34Mini-Optimization Protocols The following pre-set protocols use four or six well sets in the el

Mini-Optimization Protocols35Opt mini 96 well/ SqrTable 8 shows the parameters that vary in each well of a 96-well plate. The protocol includes the f

Gene Pulser MXcell™ System Manual | Pre-Set Protocols36Opt 96 well/ Sqr, NP, DTable 10 shows the parameters that vary in each well of a 96-well plate.

Well Set Protocols37Well Set ProtocolsWell set protocols include parameters that vary in each well of a 96-well plate. The protocol includes the foll

Gene Pulser MXcell™ System Manual | Pre-Set Protocols38Mixed ProtocolsThe following protocols include a mix of protocols with a variety of well sets,

Mixed Protocols39Opt 12 well/ Exp,SqrTable 14 shows the parameters that vary in each well for a 12-well plate. The protocol varies in a wide range of

Gene Pulser MXcell™ System Manual | Pre-Set Protocols40Uniform 12 well/ Exp,SqrThese are parameters that vary in each well for a 12-well plate. Except

Gene Pulser MXcell™ System Manual | Operating Conditions ivMechanical HazardsThe Gene Pulser MXcell system contains a patented arc-protection circuit

Gene Pulser MXcell™ System Manual416 Factors Affecting ElectroporationThe Gene Pulser MXcell™ electroporation system can help you design successful

Gene Pulser MXcell™ System Manual | Factors Affecting Electroporation42Time Constant and Number of PulsesWhile square waves do not report a time const

Factors Affecting Electroporation43Although transformation of most cell types has been accomplished using plasmid DNA isolated by a variety of metho

Gene Pulser MXcell™ System Manual | Factors Affecting Electroporation44Gene Pulser electroporation buffer is universal and can be used with most cell

Electroporation Theory45Electroporation TheoryElectroporation is a physical process in which cells are exposed to a high-voltage electric field resul

Gene Pulser MXcell™ System Manual | Factors Affecting Electroporation46By changing the capacitor of the instrument or by changing the resistance of th

Electroporation Theory47Table 17 lists the fractional voltage decrease (% droop) associated with pulse length at various sample resistances for the h

Gene Pulser MXcell™ System Manual | Factors Affecting Electroporation48Resistor Pulse ModulationThe Gene Pulser MXcell electroporation system uses a n

Gene Pulser MXcell™ System Manual49Appendix A: PulseTrac™ SystemThe Gene Pulser MXcell™ electroporation system uses the PulseTrac waveform delivery

Gene Pulser MXcell™ System Manual | Appendix A: PulseTrac™ System50PulseTrac Diagnostic Algorithm The PulseTrac diagnostic algorithm tests and selects

vWarrantyThe Gene Pulser MXcell electroporation system is warranted against defects in materials and workmanship for 1 year from date of purchase. If

Gene Pulser MXcell™ System Manual51Appendix B: TroubleshootingFollow these troubleshooting options:1. Little or no transfection or cell viability.•

Gene Pulser MXcell™ System Manual | Appendix B: Troubleshooting52

Gene Pulser MXcell™ System Manual53Appendix C: ReferencesAnderson, M.L.M., Spandidos, D.A., and Coggins, J.R., Electroporation of lymphoid cells: fa

Gene Pulser MXcell™ System Manual | Appendix C: References54

Gene Pulser MXcell™ System Manual55Appendix D: Product Specifications and InformationThe following tables provide product specifications and informa

Gene Pulser MXcell™ System Manual | Appendix D: Product Specifications and Information56Product InformationTable 20 lists catalog numbers and descript

Life Science Group 07-0545 0807 Sig 110610010739 US/EG Rev BBio-Rad Laboratories, Inc. Web site www.bio-rad.com USA 800 4BIORAD Austral

Gene Pulser MXcell™ System Manual | Warranty vi

Gene Pulser MXcell™ System ManualviiTable of ContentsChapter 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Gene Pulser MXcell™ System Manual | Table of Contents viiiChapter 6. Factors Affecting Electroporation . . . . . . . . . . . . . . . . . . . . . . 41