Gene pulser xcell electroporation system (English manual)

Gene pulser xcell electroporation system (English manual)

1. mm

Gene Pulser Xcell
Electroporation System

Instruction Manual

Catalog Numbers

165-2660, 165-2661, 165-2662, 165-2666,
165-2667, and 165-2668

2. Warranty

Model : Gene Pulser Xcell

Catalog No.: 165-000%

3. Table of Contents

Section 1

Section 2

Section 3

Introduction and Safety Information: The Gene Pulser Xcel!
system i a

Unpacking and System installation

4. Section 4

Section 5

Section 6

Overview of Electroporation Theory

Exponential Decay Puses

Factors Affecting Electroporation: Optimizing
Electroporation neem

E

Electroporation of Bacteral Celis

5. Section 7 :

Section 8

6. Section 9 References. .
Section 10 Specifications and Product Information

7. Section 1
The Gene Pulser Xcell™ Electroporation System

9. Warning

Section 2
Unpacking and System Installation

10. 2.2 Setting Up the System

2.2.1. Setting Up the Gene Pulser Xcell Main Unit and Connecting the ShockPod
(Cat. #2 165-2660, 165-2661, 165-262, and 165-2666)

Roar panel of Gone Pulser oil main unit showing receptaces for the power cord
nd PC Module

Fig. 2.18. Bottom panel of the Gone Pulsor cell main unit. The foot may be pulled down to raise

11. Fig. 2.2. Connecting the SheckPod tothe Gene Pulser Xcll main unit

2.2.2 Connecting the PC Module to the Gene Pulser Xcell Main Unit (Cat. #s 165-2660,
165-2662, and 165-2668)

Fig. 2.3. Rear view back panel) of PC Module showing the connection tothe Gene Pulser Xcel!

12. [2.2.3 Connecting the CE Module to the Gene Pulser Xcell Unit (Cat. fs 165-2660,
165-2661, and 165-2667)

Fig 24. Roar viow back panel)

2.24 ShockPod (Cat. fs 165-2660, 165-2661, 165-2662, and 165-2660)

14. Section 3
Gene Pulser Xcell™ Operating Instructions

3.1 Section Overview

15. 3.2 Front Panel and Home Screen

3.2.1 Description of the keypad

16. Fig. 94. Gone Pulse Xcel front panel. Sao Section 3.21 for an explanation of the key functions

3.22 Home Seroon

17. Fig. 3.2. Gene Pulser Xcol Home screens. Al menu functions may be rondiy accessed from
key. Press tho Loft and Right Arrow keys to toggle botwoon the two screens

3.2.3 Help Screens

18. 8.3 Manual Operation

3.31 Manual Operation (Quick Guide)

19. Fig. 3.2. Exponential Decay Protocol Detall Screen. This screen shows the parameters that may
be specified for exponential decay electroporation. Entering the cuvelte size ls optional and is for

Table 3.1. Capacitance and voltage rangos for the high voltage / low capacitance
circuit and the low voltage / high capacitance circuit of the Gene Pulser Xcell for
delivoring an exponential decay pulse.

Capacitance (uF) Voltage (Y)
Beams 2600 Rampe > 600

20. Table 3.2. Rangos of timo constants that may be achieved using Gene Pulser Xcell
at several sample resistances.

Sample rosistance Time constant range (msec)
Weireuit HV Circa

21. Time Constant Protocol Detail screen

Specified for exponential decay electroporation when the timo constant is designated. Entering

22. Fig. 3.5. Square Wave Protocol Deal Serean This seroon shows the paramators that may be spec
fied for square wave electroporation. Entering the cuvette size 1 optional anf or user informa

Table 3.3. Pulse length and pulse interval ranges for the high voltage / low
capacitance circuit and the low voltage / high capacitance circuit of the Gene
Pulsor Xcoll for delivering a square wave pulse.

Voltage (V) Pulse Length Pulse Interval No. Pulses (max)

24. Exponential Decay: Results Screen

pulso. The table gives the calculated Umo constant (TC) and the delivered voltage Voltage)

Time Constant: Results screen

‘The tabie gives the delivered time constant (TC) and voltage (Voltage) as walls the selected
Capacitor (Capacitance) and resistor (Resistance)

‘Square Wave: Results screen

LES

that the x-axis may not be to scale. The ‘the actual puise length (Pulso Length), the
Salivered voltage (Voltage, the calculated droap (% Droop), and, when multiple pulses are given,
{he pulso interval (toral) and the number of pulsos Number)

25. 9.3.6 Saving a program from Manual Operation

User Directory screen

Fig. 3.9. User Directory screen. This is an example ofthe fest User Directory screon. Toggle
botwoon thi seroen and tho second sereon with Usors 7-12 using the Left and Right Arrow keys

26. User Protocols screen

Fig. 3.10. User Protocols servon. This is an example of th frst User Protocols seroon. Togo
between this screen and the second screen with Protocol 7-12 using the Left and Right Arrow
Key. Won purchased, the inital Usor Directory scroon wil have only numbers without any

3.3.61 Saving a protocol in a location without a named User Protocol entry

27. Warning screen

28. 9.4.2 Electroporation using Pre-set Protocols

29. Pre-Set Protocols Screen

otocols Screen. There are Pre-set protocols for bat ral, and mam
‘malian cols, Uso this screon to make a selection from thelist.

Mammalian Pre-set Protocols Screen (screen 1)

Fig, 313. Mammalian Pre-set Protocols Screens. Protocols are provided for 12 mammalian col
on Seroon 2. d

30. Pre-set Protocol: CHO

Fig. 914. Protocol Detail Screen for CHO coll inthe Prost Protocals menu.

Table 3.4. Optimized settings found in the Pre-set protocols function of the Gene
Pulser Xcel.

Preset
Protocol PL © pc Cuvete Cell voi
Designation Calls Pulse type (msee)(uF) (ohm) Y (em) (ul)

31. 9.4.3 Modifying Pro-set Protocol Parameters,

32. 3.5 User Protocols

34. User Protocols - User Directory Screen (screen 1)

User Protocols - User Directory Screen (screen 2)

Fig. 3.15. User Directory seroon.

User Protocols Screen

35. Select Method Screen

Fig. 3.17, Solect Method sere.

Fig. 3.19, Timo Constant Protocol Detail sereen

37. 3.5.5 Deleting a User Name and a User Protocol

38. Warning screen: delete protocol

Fig. 3.22. Waring screen: delete protocol.

39. Warning screen: delete name

3.5.6 Renaming a User Name or a User Protocol name

Warning screen: change name

40. 3.6 Last Pulse

41. Exponential Decay Protocol Optimize Screen

‘Time Constant Protocol Optimize Screen

Square Wave Protocol Optimize Screen

42. Fig, 8.28. Data Management screen

44. Data screen (User Protocol, Square Wave)

Fig. 3.90, Examples of data screens.

45. 3.9 Measurements

3.9.1 Sample Resistance Measurements

46. 9.922 Calibration and Measurement of Capacitors in the CE Modulo

47. 3.10 User Preferences

310.1 Setting the Clock

48. Fig. 3.98, LCD Brightness scroon,

to select. The LOD display ret ser Preferences screen,

3.40.3 Sleep Function Setting

50. 9.11.2 Pulso Trac Diagnostic Algorithm

The Pu Trac ts
Extender mx

Section 4
Overview of Electroporation Theory

51. 4.1 Exponential Decay Pulses

53. [able 4.1 Droop associated with pulso length at various sample resistances for the |
high-voltage and low-voltage ranges for the Gene Pulser Xcell

Droop %

Sample
Resistance
(ohms)
20
200
1000
3500

Fig. 4.1. (A) Exponential decay pulse from a capacitance lscharge system. When a capacitor,
charged to an inital voltage Vo, a ischarged into calls, the voltage applied to tho cols decreases
Vo 09. Inthe special caso whore t= CR then Ve. The value CR is known asthe time con-
stant ofthe voltage decay. The shorter the time constant the faster the decay, (8) Square wave
pulse from a capacitance discharge system. The puise length i the timo the calls aro subjected
{o the ischarge. During the pulse the voltage again decreases in an exponential decay so that at

54. ‘Section 5
Factors Affecting Electroporation: Optimizing Electroporation

57. á

tof solution a room tomporaturo.

58. Table 5.1. Resistance of Water in 0.2 cm Cuvettes to which TE has been added

Electroporation

(at worw.bio-rad.com,

6.1 E. col

6.1.1 Preparation of Electrocompetent Cells

IF: PC

62. 6.3 Agrobacterium tumefaciens.

6.3.1 Preparation of Electrocompetent Cells

63. 6.4 Bacillus cereus

6.41 Preparation of Electrocompetent Cells

64. 6.5 Pseudomonas aeruginosa

6.51 Preparation of Electrocompetent Col

65. 6.5.2 Solutions and Reagents

66. 6.6 Streptococcus pyogenes

6.6.1 Preparation of Electrocompetent Cells

67. 6.7 Lactobacillus plantarum

6.7.1 Preparation of Electrocompetent Cells

68. Section 7
Electroporation of Fungal Cells

7.1 Saccharomyces cerevisiae

7.14. Preparation of electrocompetent cells

70. 7.2 Schizosaccharomyces pombe

7.24 Preparation of Electrocompetent Cells

71. 7.2.2 Electroporation

7.8 Pichia pastoris

7.2.4 Preparation of Electrocompetent Cells

73. 7.4 Candida albicans

7.4.4 Preparation of Electrocompetent Cells,

74. 7.8 Dictyostelium discoideum

7.54. Preparation of Electrocompetent Celis

76. Section 8
Electroporation of Mammali:

8.1 Preparation of Electrocompetent Cells

8.1.4 Attached Colla

77. Table 8.1 Suggested starting cell concentrations and volumes for
electroporating mammalian cells

Cuvete Cell concentration Cell volume — Growth conditions following electroporation
(em) (cells), mí)

78. ‘Section 9
References

81. Section 10
Specifications and Product Information

10.1 System Specifications

165-2660 Gone Pulsor Xeoll Total System

165-2661 Gone Pulser Xcoll Eukaryotic System

165-2662 Gone Pulser Xcoll Microbial System

82. 10.2 Product Information

Catalog
Number Product Description