Gene Pulser XcellTM エレクトロポレーションシステム 取扱説明書
Contents
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6. 5 3100
7. Gene Pulse
8. Square wave protocol Square Wave Protocol Detail 000 3200000000000 10000000
9. 1000000000 Chaffin amp Rubens 1998 Gene Pulser 0 O 25 PC 4000 V 2 0kV 10000 Gene Pulser Xcel main unit PC module 00000 L pl
10. Home 000 32100 0000 000 0 000 0000 Time constant protocol 7000000 ENTER Time Constant Protocol Detail 0 0 0 34000000000 1000000000000 Gene Pulser 000000000 LU UL
11. Gene Pulser X cell 10000000000 144 12 x 12 100000 00 000 00 0 000 00000 5 3321 3330 3341 00000 34900000 8 3530 00 0 00 354 00000 1008 3540000 351 0000000 100000 00000 Home DD 0d D
12. P aeruginosa PAO1 pUCP19 00000 1 10 transformants 90 100000 10000 Gene Pulser O 25 PC 2000 V 25kV Gene Pulser X cell main PC module P aeruginosa 12570 0 0 25ml LBO 0 0 0 37901
13. 641 0000000000000000 Silo Suh et al 19949 000 cereus UW85 pAD1230 0000000000000000 0 2 10transformantsu 90 00000000 1000 Gene Pulser Xcell 0 0 C 50u PC 2000 1 0 Gene Pulser X cell main unit PC module 00000 B careus 0 0 0 0 125ml 100 10ml LBO 0 0 0 0 28 300rp
14. 00000000000000000000000 Delete ENTER 100 10000000000000000000000 0 0000000000 1000000000 Gene Pulser Xcell 0000 100000 100000000000000 PULSE 00 0 000000 10000 100 Protocol Results 00 Protocol Results 0 0 Protocol Detail Protocol Detail PULSE 00000 10
15. 321 000000000000000 000000000000 Gene Pulser 0 0000000000000000000 0 0 00000000000000000 LO d OU ULL 10000000 0000 100 0 000000000000000000000000000000 000 000000000 0 00000000000000000 Gene Pulser X cell 0 Protocol Directory 0 0 00 0 000000 000000 100 0 0 000000 0
16. 100000000 8 000000 159 00000000 551 60 C 1000000000000 100mg 0 0 00 12 150 00000 SOB 20g Tryptone peptone 059 Y east extract 02ml 5M NaCl 025ml 1M 90ml 000000 70 100ml 000 1000000 IM 1 0770 1M Mosop 000060 10 SOCH 100mi 5080 000 0000 0 0 000 00 000 0 6 6 Streptococcus 5 661 IO Ut
17. Prentice 1991 0000000 S pombe CHP408 pA polinario et 19930 00 0 00 00 000 000 0 0 003x 10 ransformants u g 00100 Gene Pulser X cell C 25u PC 2000 V 23 Gene Pulser X cell main unit PC module S pombe 287 Fernback 0 000 200ml ODeoo 0110 S pombe 30 1002 100 30 C 250rpm 0 0000000000 2 10 ells ml 041 05100 20 00000000 15 IUU 250 4q 4000x g 5 10 50ml 12 100000000000 225
18. Home ENTER Preset 100 Bacterial 0 0 0 Fungal Mammalian 0 0 0 100000 100000000000000000000 0000000 1000000000000000 100 ENTER Protocol Detail 0 00000000 PULSE 00000000000000000000000 BACK 1 0 Protocol Detail 1000 342 0000 000000000000000000 1000000 19900000086 121 0 100000000000 000000000 000000000 1000 00000000000 10000 Home 100 Preset Protocols ENTERO Preset Protocols 0 320000 1000 00 00 10000000000 Bacterian 0 0 00 Funga
19. 711 0000000000000000 100000 Becker amp Guarante 1991 Ausubel et al 1987 100000000 S cerevisiae 53400 3520 00000000000000 5x 1otransformants u 90 0000000 10000000 Gene Pulser X cell 02cm 1000000 25 2000 V 15kV 0 4cm 25 2000 V 2 5kV 00000 0 Gene Pulser Xcell main unit PC module OO O00 S cerevisiad 282 Fernback 500ml Y PD 0 00 S cerevisiae 2000000000 30 C 250rpm 1 10cells ODeo 0 3 0350 10100000000
20. pH 70000 1000 0 00000000000000000 10 0 0 0 0 1M 10ml 0 0 IM 9504 00000 10 SOCH 100ml 5080 2 00 0 0 B 0 0000 00000 0 6 2 Staphylococcus aureus 621 0000000000000000 1000 Lee 19950 0000000000000 Saureus RN4220 1500 00000000000000 000 1 10transformants u 90 000000000 100 Gene Pulser Xcell O 25 1000 V 229kV Gene Pulser X cell main unit 0 PC module 00000 S aureus 17x 100 0 0 0 0 3ml B2 broth 37 C 250rpm LU 150m B2 brothr 37 C 250
21. 100000 00000000 000000000000000 0000000000000 000000000 0000000000000000000000000000000 000 00000000000000000 3 3 3 31 00000000000000 Homer ENTER 0000000 00000 0 0 0 000000000 I ENTER 100 Gene Pulser Xcel 00000000000000000000000 Protocol Detail 0 0 0000000000000 3 32 000000000000000000 100000000000000000000000000 8 4 00
22. 21 C 125rpm 241 1000000000000 16 2 0 0 0 0 000 0000 7 10 000000 21 125 0 0 LU HB DULU 2 50ml 100150 10000 00000 400x 90 5 10000000 1 100000000000000000 10 50 400x 91 50 000000000 100000000000000000000000 1 19 000000 1000000000000000000000000000000000 752 0000 LU DL UO UU 504 99 00000000 15 00000000000000 8004 Gene Pulser Pre set Protocols 1000 Fungal Protocols 00000 D discoideum Protocol Detail 0 8 341 DNA I EH
23. 2 0000000000 10 0 00001 0000000000000000000000 17 100mm 100 10 100 10000000000000000000000 5 0 0 0 0 o2aemi 00000000 15kV 0 10000 30901 480 721 0000000 713 Y PD 10g yeast extract 2090 peptone 900m0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 10ml 1021110000000 1M 18220 100 8000 0000000000 12 100000 10000 20 2090 100m0 0000000 0 224 100000000 721 00000000000
24. DNA IUUUUUUU O 0000000000000 0 ShockPod 5 0 0 0 0 00 1mp 0000 12 0 00 10000000000 100000 17x 100 000000000 juu dit 1000000 5 0 2 0 401 6000 000000 30 6 00000 7 2 3 10gr yeast extract 290 0 00 000 900mg 0000000000000000 190m 20 LOU UU 0 12 0000000 21860 0000000 7000 0 00 000 0000 120 0 100 100 0 EMM 123590 Dextrose 900ml 00000000200120 0 450 0 10 090 00000 0 300 0 550 50770 20 100 60000 0
25. 3 3 6A User Protocols 00032 00000 000 00 0000 0 0000 0 00000 3100 000000000 0 2 0 0 0 0000 0 0000 ENTER 1000000000000000000 00000 00000000000000000 SHIFTO OO 000 0 0 0 000000000000014000000000 000000000000000000000000000000000000000000000 1000 Save ENTER 100 00001 ENTER Protocol Detail 0 PULSE OO00000000000000 100000000000 3 3 6 User Protocols 100 00000 00000 00000 00 000 00000 00000 1000000000000 0000000000000 000000000 ENTER 10000000000000000000 32110 00 00000 100000000000000 you want to overwrite the protocol YES NO Press BACK or ENTER to return to the previous screen Press the LEFT arrow then ENTER to overwrite the name 0311 000000000000000 NO 1000 ENTER Protocols YES
26. 2x MRS 1000000 109 Proteose Peptone Difco 211693 1 10g Beef Extract Difco 32115200 5g Y east Extract Difco 212750 1ml Tween 801 115 ml 1M 2 4 12 3 1199 00000 08 1M 9504 2250 1M MnSO4 0800000 000000 1000000000 10 1x MRS 500ml 2 MRS 200ml 10 0 0 0 30ml l 100000000 1 0 0 0 01200 7 590 Bacto 150m0 000000000 55 0000000 250ml 2x MRS 100mig 1094 100000000000000 560 075M 00000000 1241 EI 00 0 MRS 0 25070 2 5 50ml 20 000 0000000000000 150m0 0000 25 3370 2M0 0 00 0 0 2 15 090 100ml 100000000 25 0000000 227 890 00000000 250 0 000000 500ml 10000000 100000000 30 PEG 1000 15001 PEG 10000 0 0 0 200ml 00000 5990 0000 100 5
27. Gene Pulser 3000 100000000000000000000000000 10000000000000000 400 10000000000000000000000 0000000000000000 000 000000000000000000000000 0000000000000000 000 000000000000000000000000 10000 PULSED 00000000 0000000000000 PULSED 0 0000 Pulsing 000000 1000000000 1000000 000000000 10000 ShockPod 00000 100000000000000 1000000 1000 1000000000 100000000000000000 1000000 0000000000000000000000000000000000 100000000000000 0000000000000000 13 0000000000000 Gene Pulser Xcell 10000000000000000000000000000000 1000000000000 10000000000000000000 104000 100 0 E ShockP od 000 000000000000000000000000000 1000000000000 000 000000000000000000000000000 100000000 10000000 14 0000000000000 100000000000000000 10000000000000000000000000000000000000000000000000 0000000000000 10000000000000
28. 10000000000000000000000000000000000000000000000000 0 0000000000000000000000000000 0000000000000000 0 0 000 0000000000000000000000000000000000000000 0000000000000000000000000 Chang et 1992 0 Gene Pulser Xcell 1004000000000000000000000000000000 main unit CE module module O 000000000000000000000 S 2 PULSE 0000000 main unit module 0100000000000 0000 1000000000000000 10000000000000 000000 5 0 0000000000000000000000000000000000000000 00000000000000000000 0000000000 E V d 000000009 0 1000000000000000000000000000000000000000000 1000000000000000000000000000000000000000000 1000000000000000000 1995 Neumann et 20000 0000000002 00 00000000000000000000000 0 000000000000000000000000000000000000000
29. x 100000 CE module 00000 000000 0000 0000000000000000000 0000 1000000000000000 Delete Clear 0 0 0 00000000 000 10000 00 000 00000000000 Gene Pulser X cell 0 PULSE 00000 00000000 PULSEJIJU D Pulsing O00000 Protocol Results 00000000000 5 33511 3870000 Protocol Results O Protocol Detail 0000000000000000 Protocol Detail 10000000 1000 10000000000000000000 ENTER 00 000 000 0000 10000000000003 33600 00000 10000000000000000000000 38 335 Results 10000 Protocol 10 3 60 3 0 0 00 00 0 0 0000 00 00 0 0000 0000 000 0 00 0 0 00 0 0 000 00 00 Data Management
30. 100000000000000000000000000000 000000000000 Number of pulses 10000000000000 0 10000 pulse intervalg 0 10000000000000000000000000000 10000000000 10000000 0000008 42100 00000 0 100000000000000000000000 100000000000 1000000000000000000000 330 100000000000 00 00000000000000000000 0 0000000 3 3 00000000000000000 1000 10000 100000 100 500 0 050 100ms 0 19 105 10 2000 3000 0 050 5ms 50305 2
31. 1000 2 0 000000000000000000000000000000000000 0000000000000 240000 User Preferences 0 0 0 3342 000000000000000000000 0000000 ENTERO 000000 Set 0 0 33900000000 Set Clock Set clock PREFERENCE MM DD YY DD MM YY DATE XX XX XX XX XX XX TIME XX XX 3 35 Set Clock 10000 MM DD YY DD MM YYT 0000000 0000000000000000 2 00000000000 00000000000000 100000000000000 00000000000000000 0 1000 Delete Clear 00000 100 10000000000000 0 Save 1000000000 100000000 24 00000 Save 000000 User Preferences 100000 3 10 2 User Preferences DOOOOOOOO00000000000000 Screen intensity ENTERO 000000 LCD Brightness 0 3360 00000000 LCD Brightness j LCD BRIGHTNESS Press UP DOWN arrow keys to change the brightness of the LCD display Press ENTER to select 3 36 LCD Brightness 0 10000000000000000000 User P
32. 00 000000000 0000000000000000000000000 1000 100 10000000000000000000000000000000 1000 000 1000000000000000000000000000000 0 0 00000000000000000000000000 Exponential protocol Time constant protocol 0 000000000000000000000000 Square protocol Pre set protocols
33. 1000000000 30000 0000000000000000 0000000000000 0 000000000000000000 0000000000000 1000000000000 1000000000 10000000000000000 10 15 1000 100 15 00000000 00000000000000 10000000 1 MgCI4 00 100000000000000000000000 00 1000000000 1000 9 00000000000000000000000000 Pseudomonas aeruginosa 200000000 10000000000000 Haque amp 19740 10000000000000 B 00000000000000000000000000000000000 10000 OOOCOO0O0CO0O0C0C000000000000000000000 10000 100000000000000000000000000000 000 100000000000000000 10000 10 100000 100000000000000 1000000000 5 510 0 10mM TrispH80 EDTAQGOOOOOOO0000000000000000000 000000000 1000000 10000000000000000 100000000000000 1000000 DNAD 1000000000000000 100000000 100000000 0000 cu GP Xcell Instruction Manual gt T S 5
34. 0000000000000000000000000000000 Protocol Detail LO HL E 000000000000000000000000000000 00000 100000 0000 Delete Clear JUDDDDDBDUDUDBDUDUENTERJUDDUDUDUDUDUUDUUNDU 00000 10000000000000 00 10000000 Gene Pulser X cell PULSE LH D DB D PPUuLSEr utt Pulsing 000 100 Protocol Results 0000 Protocol Detail 0000 1000000000 Protocol Results 000000000 Protocol Detail E H HB B B BE UE DH 000 00 00 IO 0 ENTER 0 0 0 0 0 0 0 0 0 00000000 18 38 000 3 4 4 0000000000000000 1000000000000000000000000000 0000 5 3430 000000000 Protocol Detail 000000000 saver 0 0 0000 00 User Directory 3970000 Choose location for protocol 100 User Directory O00 0000 000000 10000000000 000000 User Protocols WwW Choose location for protocol
35. 15 209011 3000x 90 5 0000 10000000 125m 5mM 10mM 20 200000600 3000x g 5 100000 125ml 1000 3000x g 5100000000 D 100000 125ml 1000 00 3000x g 5100000000 DI 10 100000 35ml 000 1M 1000000000000000000 Oakridge 100000 G 3000x g0 51 11 000000000 0570 00001 0000000 000 12 1000000000000 10000000000 7 4 2 00000000000 100000000000000 0000000 1 0 1 0 000 0000 17 000 100000 100000002 1000 0 10 15ml 0000000 400 10000000000 DNA 5 1000 Gene Pulser Xcell Preset 5 Fungal Protocols 10000 albicans Protocol Detail 10000 5 34 DNA 1000000000000 10000000000 ShockPod ShockPod 0 HOH 00 00 PULSEG OOO 1000000000 10 1ml 00001 0000 10000 10000000000 17x 100 10000 10000000000000 100 5 15 0 0000000 1000 1 1M 100000000000 30 40 600 0 0 0000 743 0000 PD uridine 1090 yeast extract 2090 900MIJ 000000000000000 100ml 20 10000 Im 25mg YPD uridine
36. User Directory 100000 Delete 100000 1000000000000 10000000 00000 0 0323000000000 0000 ENTER 1000 User Directory 00000000 10000 YES 0 0 00 00000000 00 User Directory 1 00000000 0000000000 All User Protocols under this name must be deleted before this user name can be deleted 10000000000000 0 000 1000000000000 Do you want to delete the user protocol YES NO Press BACK or ENTER to return to the previous screen Press the LEFT arrow then ENTER to delete the name 03 23 0000000000000 35 6 0 0 00000000 00000000 00000000000000 00000000 User Directory User Protocols Clear 00000000000000000000 3242 000000000000000 0000000 User Directory 0000 User Protocols 100000000 YES 0000 ENTER 100000000 100000000 1000000000000000000000000190 2 0 000000 Do you want to delete the user protocol
37. 1000000 10 5x 10cellsymig 00000 100 100000 00000000 Gene Pulser 0O 0000 0000000000000009 00000000000000 82 00000000000 Gene Pulser 0000000 0000000000000000000000000 000 Pre set Protocols Mammalian Preset Protocols 0 0 0 100000 Protocol Detail 340000000000 0000 00000000 000000000000000 101 590 g mili 000 1000000000 10000 1008000 10000000000 1000 00000000 0 0 0 0 ShockPod 000000 0000000 0 0000000000000 00000000000000 10000000000000000 1000000 0000000 241 48 1000000000000000 59 GP Xcell Instruction Manual 181 00000000000000000 10000 IUUUUU 0000000000000000
38. Protocol Detail Save 1000000000 10000000 1000 10000000000000 321 1 00000 0000000000 10000 OO Protocol Detail HENTERII LH D 0 User Directory 3991000000000 100 10000000000 8 334 100000 10000 10000000000 1000000000000000000000000000000 user Protocols 0 0 a 0 0 0 D D D 10000000000000 0 Do you want to overwrite the protocol YES NO Press BACK or ENTER to return to the previous screen Press the LEFT arrow then ENTER to overwrite the name 35 5 00 00000 000000000200 00000000000000000000 1000000 1000 Home 0 3 20 User Protocols 0000000 User Protocols 0000000 User Directory 3159900000000000000 User Directory O00 0000000000 000000 10000000 0 ENTER User Protocols 100 10000000 1000000 Delete 10000000000000000 NO 000000 000 0000 0 ENETRO 00000000 User Protocols 10 1000000000000 YES 0000 1000000 10000000000000000000 Use Protocols
39. ShockPod 000000000000000000000000000000000000000000000 0000000000000000000000000000000000000000000000000 000000000 000000000000000005 0 00000000000005 39400005 0000000 0000000000000000000000000000000 0000000000000 000000000000000000000000000000000000000000000 0000000000 X ge t B 00000000000 lt 00000 LA sA Cir MT ART 02 5 ShockPod 00000 0 000000000000000000000000000000000000000000000000 O ShockPod Gene Pulser X cell main unit 000000 000000000000 0 00000000000000000000000000000000000000 2500000000 0000000000000 E 000000000 2 6 Gene Pulser ShockP od 000 GP Xcell Instruction Manual 6 030 0000 3 1 Gene Pulser Xcelll 0 0000000000000000000000 10000000
40. 10000000000000000 Gene Pulser Xcell Pulse 00000000 Pulse 000000000000 10000000000 100000000 10000000000000000000000 100000000 10000000000000 0000000000000 19 100 00000000000 00000000000 10000000000000000 1000000 10000000000000000 3 11 1 Pulse Trac Pulse 00000000000000000 100000000000000 00000000000 1000000000000000000000 0 Pulse Trach 1000 0 0000000 Pulse 0000000000000000000 C LI LOU UO OU EI D U 100000
41. 3 2 User Protocols 68000000 User Protocols User Directory 0 3 15 1000000 User Directory 1000000000000000000000 1000 ENTER User Protocols 131610 0000000 1000000000000000000000000000000000 1008 355 0 000000000000000 0 0 0 0 0 Select Method 10 3 1 1000 10000 Exponential protocol 1 0 00 0000 000 T ime constant protocol 10000 Square wave protocol HL 0 0 0 0 0 000 0 Protocol Detail 0 00000000 User Directory USER DIRECTORY Labguy Malcom Smith Wesson Oo 0140720 User Directory 000020 USER DIRECTORY O 03 15 UserDirectory User Protocols USER PROTOCOLS Userl Monkey Human 01450120 0316 User Protocols 23 GP Xcell Instruction Manual Select Method SELECT METHOD l Exponential protocol 2 constant protocol 5 wave protocol 3 17 Select Method 0 Exponential protocol
42. ebd 53 Saccharomyces cerevisiae 66 666 53 VAS EG s 53 1121 GAO ee u Tasu 53 TT eet asna DT 54 Schizosacchar OMy CES POMD Es u Eco QO uu E e P PC RN CE ERE EE E EE S Y 54 PC UR 54 22 2 apap 54 72 31 55 Pichia pastori S sect pb 55 794 EVE EE aa aan 55 732 EO EV 56 tities Meese e hee h uu AR RE I 56 Candida albicans pie u S as Rapin to ey eta Ae 56 741 SET GE E LEE ob oe mele lye cote 56 742 EE GG aaa ass asa a doles o pagu nr eh 57 143 EWI ee E Sa eis 57 Dictyostelium discoideum CR ERE a 57 751 S ap LETT 57 752 VEE DE ua a e t 58 fists EP ET PEEL a
43. 00 000 0000000000000000000 100 1000000 Dower et 19921 Trevors et al 1992 100000000 coli 1000000000000000 1990 LU UH ULLLL Scerevisiad LU 000000 60 10000 Becker Guarente 1991 100 D D LU Anderson et al 1991 5 2 DNA 0000000000000 10000000 100 10000000000000000000 000000000000000000000 00000000000000000 10000000 100 0 Barsoum 1995 Candida Thompson et al 1998 Pichia Cregg amp Russell 1998 T etrahymena Gaertig Gorovsky 19950 0000000 DNA 10000000000000000000
44. User protocols 000000000 14 0 000 0 0 00000 0 0 00 0 0 0 0 00 00 10000000000000000 Last pulse 100000000000000000000 Optimize 1000000000000000000000000000000 Data 100 000 00 00 000 00 Measurement Gene Pulser Xcell main unit CE module 1000000000 1000000000000000000000000 148 User 5 1000000000000000000000000000000 9 GP Xcell Instruction Manual 0 1000000000000000 1000 1000 Exponential protocol Time constant protocol Square wave protocol Pre set protocols User protocols HOME 6 Last pulse 7 Optimize 8 Data management 9 Measurements 10 User preferences 03 2 Gene Pulser 000000000000000000000000000000000 0000 000000000000000000000000000000000000000 3 2 3
45. 6 31 0000000000000000 Lee 19957 1 0 0 Gene Pulser Xcell 00 25 FO 2000 V 224kV Gene Pulser X cell main unit PC module 00000 A tumefaciens 2 81 30 C 00000000000 50 10x 10 cells ODssd 1 00 0000000 500ml 3000x g 1d 000000000 10 10700000000 100 0 000000 500ml 3000x g 191 00 000 000 00 0000 000 100000 U LI 1000000000 10 25 30ml Oakridge 100 4 3000x gf 10 10 05 10000 100000000 15ml 5x 10 cells 0 0 0 000 D 000 10 00000000600 100 15ml 2 709100000000 0 0000000 6 32 00000000000 1000 100ng Su 00000 15 0000000000000000 10000000000 00000000000 204 0 0000000000000 000 0000 00000 17x 000000000 1ml LU D IU UL 000 Gene Pulser Home Preset Protocols 0 B 0 000 0 Bacterial Protocols A tumefaciens Protocol Detail 1008 34000000000 0 0 DNA 1000000000 0000000000000000 LU UO UU 100000 ShockPod ShockPod 0 p PULSEG 0000 ShockPod 000000000 170 00000 17x 000000 100000 0 00 000000000 10000 5msec 24VO 00000 30 C 2
46. 25 GP Xcell Instruction Manual
47. 10000000 Exponential Decay Protocol Detail 9 3280 Exponential Decay Protocol Detail Labguy Protocol Voltage XXXX Capacitance XXXX Resistance ohm XXXX Cuvette mm X 3 18 Exponential Decay Protocol Detail Time constant protocol 0 0 0 0 0 0 Time Constant Protocol Detail 0 0 3190 0 1000000000000000000000 0 Time Constant Protocol Detail Labguy Protocol Voltage XXXX Time constant mseo XXXX X Cuvette mm x 3 19 Time Constant Protocol Detail GP Xcell Instruction Manual 24
48. 100000000000000 GP Xcell Instruction Manual 58 080 000000000000000000 Gene Pulser Xcell main unit CE module 0 0 0 81 0000000000000000 8 1 1 00000000 10010 2000000080080000 demo 0000000000 500 700 confluent 0 0000000000 00000020 10x 10 ells flakg 00000000001 10000000000
49. cells ml 029 1x10 100 48 well plate with 0 5 ml growth media 0 20 5x10 2009 6 well plate with 2 growth media 0 4 2 5x10 400 6 well plate with 2 ml growth media 8 3 00000 109 0 0 0 Invitrogen Trypsin EDT A 0 05 trypsin 053mM EDTA 5 0 0 0 0 Invitrogen Phosphate buffered saline 137mM NaCI 27mMQ KCI 9 5 00000000 pH73 000 100000 HEPES buffered saline 10mM 5 pH7 3 140mM NaCI 00000000000 Phosphate buffered sucrose 272 pH7 3 0 1000 0 0 HEPES buffered sucrose 2727 0 0 00 0 0 10mM HEPES 730 0000000000000 GP Xcell Instruction Manual 60 090 0000 Anderson M L M Spandidos D A and Coggins J R Electroporation of lymphoid cells factors affecting the efficiency of transfection J Biochem Biophys Meth 22 207 1991 Andreason G L and Evans G A Optimization of electroporation for transfection of mammalian cell lines Anal Biochem 180 279 1989 Apolinario E Nocero M Jin M and Hoffman C S Cloning and manipulation of the Schizosaccharomyces pombe his7 gene as a new selectable marker for molecular genetic studies Curr Genet 24 491 1993 Ausubel F M Brent R
50. GP Xcell Instruction Manual 18 Pre set Protocols PRE SET PROTOCOLS l Bacterial 2 Fungal Mammalian 3 12 Pre set Protocols 000000000000000000000000000000 000000000000000000000000000 Pre set Protocols 000010 1000 PRE SET PROTOCOLS MAMMALIAN CHO 958 BHK21 Wwe Pre set Protocols 00020 PRE SET PROTOCOLS MAMMALIAN 549 CV1 K562 HL60 Jurrkat HuT 78 03 13 Protocols 00 1200000000000000000000000 00000000000000000000000000000 00000000000000000 Protocol Detail PROTOCOLS DETAIL CHO Voltage 160 Pulse Length msec 15 00 Number of pulses 1 Pulse interval seo 00 0 Cuvette 2 P 03 14 000000000000000000 Protocol Detain 19 GP Xcell Instruction Manual 000000 000000 Mammalian 1 Mammalian 2 Mammalian 3 Mammalian 4 Mammalian 5 Mammalian 6 Mammalian 7 Mammalian 8 Mammalian 9 Mammalian 10 Mammalian 11 Mammalian 12 Bacterial 1 Bacterial 2 Bacterial 3 Bacterial 4 Bacterial 5 Bacterial 6 Bacterial 7 Bacterial 8 Bacterial 9 Fungal 1 Fungal 2 Fungal 3 Fungal 4 Fu
51. un UJ OO gt 3 100000000000000000000000008 42 Gene Pulser 103210000000000000 0 Square protocol ENTER quare Wave Protocol Detail 0 0 0 35100000000 5 7 0000 Square Wave Protocol Detail PROTOCOL DETAIL SQUARE WAVE Voltagecv X X X X Pulse len th mseo X X X X X Number of pulses 1 Pulse interval sec 00 0 Cuvettecmm x 03 5 Square Wave Protocol 000000000000000000000000000000000000000 00000000000000000000000000000 GP Xcell Instruction Manual
52. 100000 Gene Pulser X cell 25 PC 2000 V 2 0kV Gene Pulser X cell main unit PC module 00000 S pastoris 282 Fernback 0 0 0 500ml 5 pastoris 30 90000 2100000000 30 C 300 0 0 0 000000000 7 10 mil 0240 0300 0 0 10000000000 500711 10000 4 3000x gri 9000000000 10000 10070 00 0 0 YPD HEPESU 000000000 100000000 25 0 1M DTT 30 C 5 10000 1000000 1 500ml 000 4901 3000x 90 5 000 1000000 1 00000000 5070 000000000000 100 1 5 DO 0 0 00 0 00 4 3000x gf 50 0 0000000 000 1000 1M 25ml 1000 30m Oakridge 4 3000x 50 0000000000 1000 1 13m 1 107 5 ml 1000 10000000000000000000000000000000 GP Xcell Instruction Manual
53. 1000 1ml 1M 17x 100 0 0 0 0 1000 100000000002 00 100000000000 1000 DNA 91000 15ml 10000 400 Gene Pulser Xcell Preset Protocols Fungal Protocols 0 0 0 0 0 0 0 5 pastoris Protocol Detail 05 3400 DNA U 100000000000 ShockPod ShockPod 0 0 0 0 0 0 PULSE 000 1ml 1 00000000000000 0 17 100ml 100000000 0 0000000 5msec 000 20kVDp D 0 0 0 0 1000090000000 LU U tL 10000000000 0 30 q 10 00000 1 1M 0 01000000000 100 1100000000000 30 C 720 960 0 7 3 3 HU LL YPD 1090 yeast extract 2090 peptone 00 00 0 000000000000 100 10 20 0000000 Y PD HEPES 100ml Y PDO 20ml 1M 5 8 01 1M 18220 100 800 0 0000000001400000000000000 1000 20 2090 10 0000000 0227 000 U LU LI IM 5 8 0 23 890 HEPES MW 22383 65ml 5NQ 0 pH 80 1000 100ml 1000000 2 3590 0 0221 m 1000 1000 IM DTT 3 8590 DT T 22ml 1000000 25mg 100000 022u mti t HU UU 1000 74 Candida albicans 7 41 00000000000 deBacker et 1999 C albicans 10000000 100000000 400transformants u og 00000
54. 37 GP Xcell Instruction Manual 41 Gene Pulser 0 0 0 0000000000000000 10000000 Vt Vo e t RO Vditigguudddesgultimseqi III IU n e u 0 lt 0 msec 1000 1000000000000000 0000
55. 8 3275 FU LU D LU 100000000 0 0 0 1000000 0 00000000 I I 3 8 c D 8 5 1000000000000 00044 000000000000000000 00000 51 00 000 0 droop 524 0 0 0000000 droop 5 1 B 8 GP Xcell Instruction Manual 38 041 0000000 100 droopf 0 0 00000 5 10 20 0000000 0000 msec 20 0 051 01099 02239 200 0 510 1099 223 1000 2 55 5459 1129 3500 8 93 1941 39 0 Vo
56. Xcell Instruction Manual 44 Gene Pulser Xcell
57. 10000 17 341 0000000 POTA 18 342 TEE GCE OO ee i ete 18 000000 21 344 000 1000 ER HE RR 21 MES 22 351 22 352 ER ui 22 35 3 aaa aa paqa 22 354 25 355 1000 26 3 5 6 27 10000 MM PE 28 10000 T ayayay 28 gh 29 n eec 32 391 E 32 392 33 La P T RS 3101 34 3102 LITT 34 3103 dain 35 Pulse Trach E MRNA 35 CH NIME Ji Neo 35 3412 Pulse T rad FEE ED ELE 36 HOO E ENERE s 37 seve ek TD 38 ua ph paha a 38
58. Gene Pulser Xcell 100000000000 10000000000000 0 10000 3000V 1000 00000000 Gene Pulser Xcell main unit 00000 10 15 21 F 000000000 CE module 1000000000000 100 0 00 000000000 Gene Pulser X cell main unit CE module moduld Shock CE module 0000000 00 0000000000000000000000 0 module 0 0 00 100000000000000000000000000000000 module 0000000000000000000 module 0 000000000000000000 0 modules 50u F 3275y 254 1000000000000 10000 00010 10000000000 3275 F 100000 PC module 0000000 100000000 1000000000 module 0 500 10000 500 1 000000000000 100000000000 0000 000 00 0000 00 0000000000000 0 00 100 0000000000 module 00000 PC module 000000000000000 0 0 PC module module 0 main 000000000000000000 0 main unit 00000000000000000 11 000000000 0 610101 EMC 0 61326 Class Class 10000000000000 000000000000 0000000000000000000000000 10000000000000000000000000000000 00000000000000000000 0 Gene Pulser Xcell 0000
59. Gene Pulser 1 0000000000 0 0 0 0 0 00000000000000000000000000 000000000000000 0000000000000000000000000000000000000000000 100000000000 100000000000 00000000000000000000 00000000000000000000000000000000000000000000000 GP Xcell Instruction Manual 12
60. 8 322 dec 9 323 10 Ne PDC 10 331r TO HUB B BHI 10 332 JOO EEE EEE E PIER 10 333 aaa 12 a E aer oe on oe nnt rH nn 13 5335 eco a ED 14 336 One b S S Ses rr 16 336A H Hill i 17 33065 U aaa susu ter ob ab sS 17 34 35 36 3 7 38 39 3 10 3 11 040 41 42 050 51 52 53 54
61. 7 3 Pichia pastoris 731 0000000000000000 Cregg amp 5 1998 0 0000 pastoris X33 Sacl pPICZA 10 transformants Hg
62. 0 0000000 BLU EL BE LE ET LH 00000 P Gene Pulser X cell 11 GP Xcell Instruction Manual PULSEG 000000 OOOO PULSEQGQOQ000000000 Puling 0000000 100000 00000 0000 0 Protocol 1000000000000 IUUUUU 8 3350 36 LU OUI LI Protocol Results 0 Protocol o0 0000000000000 Protocol Detail 1000000 10000000 000000000 0 00000000000000000000 08 000000000 10000000000000000000000 39 18 41000 Gene Pulser X cell
63. 100 50000000 250 32750 25000 00000090 00000000090 0020 000000000 0 050 100 00000000 1010 0000000000 seco 0 1010 module 2000 30009 10000 100 250 50 00000090 500 10000 500000 gt 00000000090 2002000 25 000 6009 25000 3000 0 000 00000000 msec 0 050 5 00000000 102 0000000000 seco 5030 Xcell Instruction Manual 64 102 Ordering Information 000000 165 2660 000 Gene Pulser Xcell 0000000060 Gene Pulser Xcell main unit Gene Pulser Xcell CE module Gene Pulser Xcell PC module Gene Pulser Xcell ShockP od 0 1 0 2 0 4 50 165 2661 1 Gene Pulser Xcell CEG O00 0000 Gene Pulser Xcell main unit Gene Pulser Xcell CE module Gene Pulser Xcell ShockP od 00000000000000000 2 04cm 500 165 2662 1 Gene Pulser Xcell PCO 00 0000 Gene Pulser Xcell main unit Gene Pulser Xcell PC module Gene Pulser Xcell ShockP od 00000000000000000 1 2 0 500 165 2666 1 Gene Pulser 00000 165 2666 165 2667 165 2668 165 2669 165 2086 0000 Gene Pulser Xcell main unit Gene Pulser Xcell ShockP od Gene Pulser Xcell main unit Gene Pulser Xcell CE module Gene Pulser Xcell PC module Gene Pulser Xcell ShockP od 000000000 Co Co 000
64. 8 3520000000000 100 User Protocols 0000000000000000 000000000000000000000000000 336A 00000 1000000008 8 355 100000000 ENTER Protocol Detail 0 00 000000 PULSED 0 0000000000000 21 GP Xcell Instruction Manual 35 0000000000
65. aee DM 40 he ie cis E 40 DNA 40 10000 3E itii tee een Reiten 41 eT 42 060 Fp ETE TT BE ae ai ios 44 6 1 ai ERR 44 6 11 aa rc 44 6 12 aeo tou na aga 44 94 32 nu x 45 62 5 e b E RR 45 Lov ER 45 622 ul eb Ep S EE e Et eee Era 4 623 4 63 Agrobacterium 430 0 ense he 47 631 i adl bed 47 632 aaah EEN 47 6 33 E t huq
66. 100 100 500 4000x gr 9100000000 1000000 000000000000000000 1070 10ml Lithium Acetate OO 00 0000 0 30 C 8 45000 0 1000 25ml 1M 3091 5 15 10000 500 0 000000000 4 1 4000x 90 5 0 0 000 0 0 0 0 0 0 0 000 10 10000 0 50ml 000000 5 1000000 4000x 90 5 00000000000 11 10001 000 251 30ml 0 30ml Oakridge 0 00000000 4q 4000 5000000000 12 100 1 0 13 15 0 0 0 0 1x 10 cells 10000 100000 7 12 00000000000 AM H EH BL B D B D 00 17x 100 0 0 0 D 100000000000000 0 20 000 00000 100000000000000 DNAQ 50 100191 55 0000000 1510 000000000000000 O02cmt D 0 0 000 00 401 0 04cm 0000000 80 100 1051000000000 Gene Pulser Xcell Preset Protocols Fungal Protocols 0 0 D D 0 0 0 0 2cm 0 0000 5 cerevisiae 2mm cuvette Protocol Detail 04cm ENTERO I 0 0 S cerevisiae4mm cuvette Protocol Detail J 000000008 347000000000 DNA 10000000 100 10000000000000 00 ShockPod 00000 ShockPodj 00000 00000000 53 GP Xcell Instruction Manual
67. What data do you want to view ALL data l Reverse chronological order 2 Chronological order Data from a specific date Chronological order 4 Reverse chronological order 03 28 Data Management Date From what date do you want to view data Month MM Date DD Year 3 29 Date 55 0 00000000000 00000 0000 000000000000000 0 00000000000000000000000000 00000 H Data Management 00000 0000 Data output 00000000000000000 100 0 00000000000 100 LU GP Xcell Instruction Manual 30 Data output Data output Data output Data output User 11 Protocol 11 Settings V 1230 PL cmsec 185 00 P 12 12 3 droop 12 User Protocol S quare 12 12 01 20 80 Output 1230 123 00 12 12 3 12 Data output Manual Mode Exponential Data output Manual exponential Settings V 1230 1230 R OHM 1000 TC mseo Cuvette mm 1 12 12 01 20 21 Out
68. 10000000 10001100001 00 U UL Gene Pulser 00000 LU UU 00 0 0000 00000 10000000 0 module 1000000000 module module 0 10000000000000 00000000 3 2000 000000000000 1000000000000 100000000000000000000000 32 1000000000000 1000010001 1000 00 00 Gene Pulser Xcell UD UO D OE EL GE EE CE DE D D CE GE HD 0000 LU UU EB DE UE OG 00 1000000000000 2900000000000 1000000000000 LU PULSE H1 Hl Hl BH E E E E E E E E EL EL E 00000 0 1000000000008 39 032 00000000000 20 0 50 65 5 0 2 1 0 200 5 0 655 2 0110 0 1000 2503275 10050 000000 10 500 00000 0 0 200 0 000000 GP Xcell Instruction Manual 12 Time Constant Protocol Detail PROTOCOL DETAIL TIME CONSTANT Voltagecv X X X X Time constantCmsec XXXX X Cuvettecmm x 00000000000 00000000000 034 Time Constant Protocol Detail 00 00000000000 00000000 00000 000000000000000000
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72. 0 00000000000 Do you to delete the protocol YES NO Press BACK or ENTER to return to the previous screen Press the LEFT arrow then ENTER to delete the name 03 22 00000000000000 GP Xcell Instruction Manual 26 User Directory 00000000
73. 47 64 Cereus u 48 641 RE EET ET 48 542 DD aaa ss doles ies nl D pagu nre 48 643 sun m 48 65 Pseudomonas aeruginosa dixi e t e eet Eod Dee e HO E Or CO E CR ERE a ERE 49 551 ERE EE ET BE E SET E E o EGET Eoo x a s tnn tne FREE Ren 49 652 PESE EE itae eo Qam oe ER me 49 653 EXER EXE EORR MERE EUN ED NOR 49 66 Streptococcus PYOGENES 50 661 ENSE EEE uo t the ua a Eee tee EUER 50 62 E EL BE AE 50 66 35 EET 51 6 7 Lactobacillus plantarum 2 2 2 ee m 51 671 A 51 672 Gh 51 eee ET 52 0 70 71 72 73 74 75 080 81 82 83 O90 0100 10 1 102
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76. YES NO Press BACK or ENTER to return to the previous screen Press the LEFT arrow then ENTER to change the name 03 24 000000000 27 GP Xcell Instruction Manual 3 6 000000000 Home DOO32000000000000000000000 Last pulse DO 00000 ENTERODOOO 100000000000 Protocd 00000000000000000000000000000 100000000000000000 3 7 00000000 100000000000000 1000000000000000000000 10000000 1000000008 3321 333 00000 000 0 00000 00 0000000 Home 3200000000000 100000000 Optimize 000 Select Method 3177 0
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79. Kingston R E Moore D D Seidman J G Smith J A and Struhl K eds Current Protocols in Molecular Biology John Wiley amp Sons NY 1987 Barsoum J Stable integration of vectors at high copy number for high level expression in animal cells in Methods in Molecular Biology vol 48 Nickoloff J A ed Humana Press Totowa Nu p 225 1995 Becker D M and Guarente L High efficiency transformation of yeast by electroporation Methods Enzymol 194 182 1991 Bottger E C High efficiency generation of plasmid cDNA libraries using electro transformation BioTechniques 6 878 1988 Bringel F and Hubert J C Optimized transformation by electroporation of Lactobacillus plantarum strains with plasmid vectors Calvin N M and Hanawalt P C High efficiency transformation of bacterial cells by electroporation J Bacteriol 170 2796 1988 Chaffin D O and Rubens C E Blue white screening of recombinant plasmids in Gram positive bacteria by interruption of alkaline phosphatase gene phoZ expression Gene 219 91 1998 Chang D C Chassy B M Saunders J A and Sowers A E eds Guide to Electroporation and Electrofusion Academic Press Inc San Diego 1992 Chu G Hayakawa H and Berg P Electroporation for the efficient transfection of mammalian cells with DNA Nuc Acids Res 15 1311 1987 Cregg J M and Russell K A Transformation in Methods in Molecular Biology
80. vol 103 Higgins D R and Cregg J M eds Humana Press Totowa NJ p 27 1998 DeBacker M D Maes D Vandoninck S Logghe M Contreras R and Luyten W H M L Transformation of Candida albicans by electroporation Yeast 15 1609 1999 Dekeyser R A Claes B deRycke R M U Habets M E vanMontagu M C and Caplan A B Transient gene expression in intact and organized rice tissues The Plant Cell 2 591 1990 Dennis J J and Sokol P A Electrotransformation of Pseudomonas Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa Nu p 125 1995 Dower W J Electroporation of bacteria a general approach to genetic transformation in Genetic Engineering Principles and Methods vol 12 Plenum Publishing Corp NY p 275 1990 Dower W J Miller J F and Ragsdale C W High efficiency transformation of E coli by high voltage electroporation Nuc Acids Res 16 6127 1988 61 GP Xcell Instruction Manual Dower W J Chassy B M Trevors J T and Blaschek H P Protocols for the transformation of bacteria by electroporation in Guide to Electroporation and Electrofusion Chang D C Chassy B M Saunders J A and Sowers A E eds Academic Press Inc p 485 San Diego 1992 Drabeck J J Glasspool Malone J Somiari S King and Malone R W Cutaneous transfection immune responses to intradermal nucleic
81. 1000000000000000000 300 100 0000000000000 mg 5 17x 100mm p in Oo Oo Oo Oo Oo Oo N O 3 Gene Pulser Xcell Home Preset Protocols 0 00000000 0 Bacterial Protocols 100 S aureus Protocol Detail 1000000 5 340000 10000000 DNA 000000000000000000000000 LU 10000000 0 ShockPod ShockPod 1 0 0 00 0 0 PULSE 0000000 ShockPod 000 0000000
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83. Chu et al 1987 Showe et al 1990 GP Xcell Instruction Manual 40 1000000000000000000000000 000000000000000000 10000000000000000000000000000000000000000000 000 0 0 00 0 0 0 00 0 0 0 0 0 00000000000000000 0 0000000000000000000000000000000000000000000 0 10000000000000000000000000000000000000000000 1000000 1990
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86. Simon amp Frretti 1995 S pyogenes CS1019 pDC1230 0 00000000000 00 0 0 0 1 JOtransformantyu 90 0000000 10000000 Chaffin 5 1998 Gene Pulser Xcell O C 25u FO 2000 V 2 1kV 10000 Gene Pulser Xcell main unit PC module 00000 0 0 S pyogenes 25ml 00000 20mMr 0000000 201 0000 37 0000 18ml 500ml 20mM 250ml 0000 02 100 10 cells mig O 0 0 1210 00000000 000014 100 1 500ml 0 000 4201 6000x g 10 100 1000000000000 1070 0000 0 00 00000 00 000 25 0 0000000 8000x g 10000 00 30ml 1000 5 0 000 0000000 30ml 12000x 90 0000 0000 1000 15 LU 1000000000000000000 662 00000000000 00000000000 0059 0 54 90 00000 15 100000 100000 2004 000000 1000000000051 1900000000 2004 00000 1000000 02cm 100000 UE D UO BL 0000000 1000000000000 17x 100mmg 0000 000 1070 000 00000000 Gene Pulser Xcell Home 0 Preset Protocols 00000000 Bacterial Protocols 00000 S pyogenes Protocol Detail 1000000008 3401000000000 10000 5 ShockPod PULSE 00000000 0000000000054 00000 10000 100000000000000 98mseq 21kV 000 100000 000 100000000 10000000 10mg 0 17x 100mm 00 00000 37 C 21 100000 37G 1
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102. 10 0 65 150 0000000000 Wilson amp Gough 1988 Heery amp Dunican 1989 acobs et al 1990 0 00 0 0 0 00 Bottger 1988 Zabarovsky amp Winberg 1990 1 1 0 10000000000000000000000 53 00000000000 0000
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104. 10000000000 Thomson et al 1999J Gene Pulser X cell 0 25 PC 2000 15 00 Gene Pulser Xcel main unit 0 PC module 00000 C albicans 00000 50 YPD uridine 0 0 17x 100 0 0000000 3091 2501 0 0 000000 0 1000 28 Fernback 500777 Y CD uridine 30 C 2501 0 0 0 0 0 0 080 50 10x 10 cells mI ODso 03 045700 100000 1000000190 100 500 000 0 GP Xcell Instruction Manual 56
105. 1090 yeast extract O 2090 peptone 900ml 1590 10000000000 100 0 00 0 200 100 25mg ml uridine 182 29 0000 800 1000000 12 10000 10000 2010 2090 60ml 10000 100m0 0000000 022 100000000 5mM c 10mM 1770 1M Lithium Acetate 10 2g 10077 20ml 0 1M DT 154 g 100ml 2ml 1M Tris 7 5 0470 0 5M EDTA 17667 00 00 0 00 0 0 00 0 00 00000 2001 0 0 1000 7 5 Dictyostelium discoideum 751 0000000000000000 10 000 E Howard et 1988 Knecht amp 1995 10000000 Gene Pulser Xcell 000000 0 V 10kV 0000 0000210000000 5sec 50 7 10cells mig 0 0 0 50070 0 0 50 0 40ml 1000000000 1000000000000 Dictyosteiumg 0 21 C 0 0O 12100000000 57 GP Xcell Instruction Manual
106. 310 00000000 Measurements Measurements l Sample Resistance 2 Capacitor 03 31 Measurements 391 0000000 1100 9000000000000 59000000 000000000000000 0 11000010000 0 1100 ohms Measurements 330 0 Sample Resistance 00000 0000 Sample Resistance 3320 00000000 Sample Resistance Sample resistance Resistance ohms XXXX Place sample between electrodes 0 3 32 Sample Resistance O 0000000000000 ShockPod 0 0 0 0 000000000 0 0 0 150 12000 ENTER GP Xcell Instruction Manual 32 3 9 2 moduleQ 1g 30700 10000000 0 0 00000000000000000
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108. 9 Concentration nM B 3500 3000 2 5 500 LN E 2000 S 4 5 1500 2 1000 Me 8 500 62 0 0 2 4 6 Concentration nM NaCI 40 F1 NaCI 200 I MgCl 40 I 0 1 200 2 Po 6 1 40 i PO 6 1 200 I i t Po 7 3 40 p d amp 7 200 p HEPES 7 5 40 I c 7 5 200 11 051 00000000000 A0 BHO 000 QQ 50 D 0 0000000000 2 lt 0 000040 0000 299 00000000000000000 051 0000 Water Water 1u 5 Water 10 40u volume 0 600 000 600 000 11 200 1 850 200p volume 600 000 35 0009 87009 4 700 000000 0 2 0 0000405 0000 200 00000000 10099000 GP Xcell Instruction Manual 10000 D E co
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110. Home Preset Protocols 0 0 0 0 00 Bacterial Protocols ENTER D coli lmm cuvette Protocol Detail 100 02cm 00000000000 25 30kV 100000000000 Detal 00000000008 DNA 10000000000000 10000000000000 100 1000000 ShockPod 100 ShockPod 050000 0 PULSE 00000000 ShockPod 00000 100 1ml SOC 100000000 100 HEU 00000 Dower et 19881 000 000000000000000003 1r U 100 10 10 1100000000000 20001000000000000000 10000000 17x 100mm 37 C 225rpm 1 10000000 0000000001800 00 00 613 00000 L Broth 10g ryptone peptone 59 Y east extract 5g NaCI 100 0000000000000 L8rothp 000000 159 00000000 100000000 551 6q 0 00000 000 00000 0 0 100m 0 12 15ml 1269 100000 900 00 00 0 00 0 0 0 00 00000 1000 10mM Tris HCl 801 1mM EDTA SOB 20g Tryptone peptond 059 Y east extract 02ml 5M 025 1M 90ml 000000
111. Vo 2 852 55 o t gt 00000 5 10 20 0000 msec 3 349 7149 14 669 3349 7149 146 1679 35769 7309 585 1249 2556 Time 041 50000000000000000000000000000000000000 000000000000 9000 van v vo 90 000 000 0 t CROOOO 0 0 00 0 0 0 DD 000000000000000000000000000000000000000000000000000 00000000000000000000000000000000000000000000000000 000000000000000000000000000000000000000000000000000000 00009 000000000000000000000000000 Xcell Instruction Manual 050 00000000000000000000 Shigekawa amp Dower 1988 Change et al 1992 Nickoloff 1995ab 00 0 0 0 0 0 000 0 0 0 0 0 0 0 0000 0000 LU DD UB DE OG EL BD UG DE GE ET D DE CE CT E 10 Calvin amp Hanawalt 1988 Dower et 19880 Shillito et al 1985 Fromm et 1987 Dekeyser et al 1990 Joersbo amp Brunstedt 19919 0 Chu et al 1987 Knutson amp Y 1987 Andreason amp Evans 1989 Anderson et al 1989 1994 10 E S cerevisiael 10000000 LU UL BL BL I 0 000 10000
112. acid vaccination are significantly enhanced by in vivo electropermeabilization Mol Ther 3 249 2001 Fromm M Callis J Taylor L P and Walbot V Electroporation of DNA and RNA into plant protoplasts in Methods in Enzymology vol 153 Academic Press Inc p 351 San Diego 1987 Gaertig J and Gorovsky M A Gene transfer by electroporation in Tetrahymena in Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa NJ p 331 1995 Heery D M and Dunican L K Improved efficiency M13 cloning using electroporation Nuc Acids Res 17 8006 1989 Heiser W C Optimizing electroporation conditions for the transformation of mammalian cells in Methods in Molecular Biology vol 130 Tymms M J ed Humana Press Totowa NJ p 117 2000 Heller R Jaroszeski M Atkin Moradpour D Gilbert R Wands J and Nicolau In vivo gene electroinjection and expression in rat liver FEBS Lett 389 225 1996 Hui S W Effects of pulse length and strength on electroporation efficiency in Methods in Molecular Biology vol 48 Nickoloff J A ed Humana Press Totowa NJ p 195 1995 Howard P K Ahern K G and Firtel R A Establishment of a transient expression system for Dictyostelium discoideum Nuc Acids Res 16 2613 1988 Jacobs M Wnendt S and Stahl U High efficiency electro transformation of Escherichia coli with DNA from l
113. ain Heart Infusion O 3790 1000ml 000000 100 10000 L Broth 10g Tryptone peptone 5g Yeast extract 5g 102 0000000000000 GP Xcell Instruction Manual 6 5 Pseudomonas aeruginosa 651 0000000000000000 Dennis amp Sokol 19951 Smith amp Iglewski 1989
114. antarum 00000 0 500770 5 0 0 0 30 151 000000 1000 28 00000 500m 5 5800 00000000 ODso 005 0 00 0000 00 30 C 00 00 0000 0 30 000 0 100000000 500mg 000000 18 C 3000x 90 10 1000000000000 00 250 1000000 18 C 4000x g0 10 1000000000000 00 250 1000000 18 C 4000x gr 10 10000000000 2 30 100000000000 672 00000000000 100000000 10 24000000000 15mg 0 100000000 401 10 10000000000000000 0 40u 0 0000000000 0 02090 00000000000000000000000000 Gene Pulser Preset Protocols Bacterial Protocols 0 0 0 0 L plantarum Protocol Detail 0000 3400000 0 0 0 0 0 0 PULSE 1000 00000000000 000 30 100000 img 56000 000 00 0 00 00000 00000 17 100mm 000 0 000 0 30 30 42 000000 1000000 8 50 0000 30 qj 20 3 000000 51 GP Xcell Instruction Manual 673 0000
115. g NaOH 13 8 g maleic acid 4 NaOH 500ml 3 I 000 0000 100000 2 SMM 25 02 sodium hydrogen maleate 40 ml 01 NaOH pH 650 5 ml 1M MgCl 427908 000000000 125 000000000 4x Penassay broth 17 5 g Antibiotic Medium Becton 250ml 1000 SMMP 55ml 2x SMM 40ml 4x Penassay broth 5ml 10 w bovine albumin pH 70 10000000000000000000000 10000000000 10 0000000 12690 00000 0 90 10000000000000 10000 GP Xcell Instruction Manual 46 6 3 Agrobacterium tumefaciens
116. h 10g Tryptone peptone 5g Yeast extract 5g 102 100 49 GP Xcell Instruction Manual
117. igation mixtures Nuc Acids Res 18 1653 1990 Joersbo M and Brunstedt J Electroporation mechanism and transient expression stable transformation and biological effects in plant protoplasts Physiol Plant 81 256 1991 Knecht D and Pang K M Electroporation of Dictyostelium discoideum in Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa NJ p 321 1995 Knutson J C and Yee D Electroporation parameters affecting transfer of DNA into mammalian cells Anal Biochem 164 44 1987 Lee J C Electrotransformation of Staphylococci in Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa NJ 209 1995 Leonardo E D and Sedivy J M A new vector for cloning large eukaryotic DNA segments in Escherichia coli Bio Technol 8 841 1990 Lin J J Electrotransformation of Agrobacterium in Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa Nu p 171 1995 Miller E M and Nickoloff J A Escherichia coli electrotransformation in Methods in Molecular Biology vol 47 Nickoloff J A ed Humana Press Totowa NJ p 105 1995 Neumann E Kakorin S and Toensing K Principles of membrane electroporation and transport of macromolecules in Methods in Molecular Medicine vol 37 Jaroszeski M J Heller R and Gilbert R eds Humana Press Totowa NJ p 1 2000 Nic
118. koloff J A ed Methods in Molecular Biology vol 47 Humana Press Totowa NJ 1995a GP Xcell Instruction Manual 62 Nickoloff J A ed Methods in Molecular Biology vol 48 Humana Press Totowa NJ 1995b Prentice H L High efficiency transformation of Schizosaccharomyces pombe by electroporation Nuc Acids Res 20 621 1991 Potter H Weir L and Leder P Enhancer dependent expression of human k immunoglobulin genes introduced into mouse pre B lymphocytes by electroporation Proc Nat Acad Sci 81 7161 1984 Shigekawa and Dower W J Electroporation of eukaryotes and prokaryotes A general approach to the introduction of macromolecules into cells Biotechniques 6 742 1988 Shillito R D M W Paszkowski J Muller M and Potrykis High efficiency direct gene transfer to plants Bio Technology 3 1099 1985 Showe M K Williams D L and Showe L C Quantitation of transient gene expression after electroporation Nuc Acids Res 20 3153 1990 Silo Suh L A Lethbridge B J Raffel 5 H Clardy J Hanelsman J Biological activities of two fungistatic antibiotics produced by Bacillus cereus UW85 Appl Environ Microbiol 60 2023 1994 Simon D and Ferretti J J Electrotransformation of Streptococcus pyogenes with plasmid and linear DNA FEMS Micobiol Lett 82 219 1991 Smith A W and Iglewski B H Transformation of Pseudomonas ae
119. l 0 0 00 ENTER 10000000000000000000000 1000000000 00 10000000000000000000000000000000000 Protocol Detail 10000 100 Pre set Protocols Mammalian 6 Preset Mammalian Protocols 0 00 3130700000000 Preset Mammalian Protocols 0000000 00 10000000 Protocol Detail 100000000000000 Pre set Mammalian Protocols 1000000 2mmr B E U D D 7 D D CHO Protocol Detail 100000000 0 PULSEGOOOO0000000 100000 Pulsing 100000000 Protocol Results 18 33500000 1005000 0 Protocol Results Protocol Detail 00000000000000000 Protocol Detail 0000 00000000000 ENTER 1000000 100005000008 34310 0800006000 00000000000 00000000 0008 38 000
120. li 20 1000000 1000 1090 0 1 10 Shigekawa amp 1988 S aureus 7 00 0 1000 1000000 Eee 1995 10000 100000 0 0000000 Chu et 198 1000000000000 1000 000000000 Potter et al 1984 GP Xcell Instruction Manual 060 00000000000000 0 000000000000000000000000000000000000000000000 0 0000000000000000000 00000000 6 1 Escherichia coli 6 1 1 1000000000 1000 Ausubel et 1987 Miller amp Nickoloff 1995
121. lser Xcell main unit 0000000000000000000000000 24400000 0 00000000000000000000000000000000000001 9000000 0000000000022800 ShockPod 00 000 Gene Pulser Xcell unit 0 0000000000000000000 000000000000000000000 2200000000000000000000000 0000000000 5 000000000000000 00000000 0000000000 Gene Pulser Xcel 00000000000 CE module 000000 0000000000000 000000 2 1A Gene Pulser Xcell main unit 000000 0000000 module module 00 000000000000 000000 ShockP od 00000 Xcell ShockP od 2 2 Gene Pulser Xcell main unit 1 00000 GP Xcell Instruction Manual tud 2 2 2 PC module Gene Pulser Xcell main unit 165 2660 19 165 2662 10 165 2668 PC module CE module 0 0 Gene Pulser Xcell unit OO 0000000000000000 module 7070000000 Gene Pulser Xcell main 1 00000000000 module Ll 00000000000 Gene Pulser Xcel main unit 0 0000000 L1 PC module Gene Pulser Xcell main unit 00000000 CE 00000000000000 0000000000000000000000000000000000000000
122. mr 00000000 2ml 282 Fernback O 500ml 29901 300rpomp O0 000000000 030 0 0 000 0 0 1x 10 lt 5 0000000000000 0 0 14 2 250ml 000000000 424 10 000x 10 0 0 0 50ml 1000000000 0 0 10 Oakridge 1000000000 424 10 000 1010 0 0 0 100000000 05mg EPO 100000000000000000000000000000 10000000000000000000000000000000000000000 6 42 0000000000 000000 0051 14 90 101 15 0 00000000000000 1004 1000000005 1900000000 1004 00000 000000020000000000000000000000000000 0000000000000 00 17x 100mm 0000020 018000000000000 Gene Pulser Xcell Home Preset Protocols O00000 Bacterial Protocols LJ LH cereus Protocol 00000 5 34 1000 ShockPod 0 0 0 ShockPod ShockPod 0000000000000 17 100mm 100000 1000000000000000000 8 6msec O00 000000000 1000 10 cr 00000 37 250rpm 10 1510 00000000 0000000000001800000000000 28 00000000 6 4 3 EPO O 05mM 05mM 272mM 000000 54470 100u IM 190 0 1M KH2PO4 810 0 1M gt 4 Milli Q 200m DOH OO 0000 0 PHO J 07 000000000000000 49100000 Br
123. mtn 10 4000x gf 5 200770 12 00000000 10ml 1 2M 0 0000 Oakridge 00 0 00000 0 15 20ml 1 2M 0000000 Oakridge 00000 4 1 4000x 57 0000000 1000000 1000000 12 0 00000000000 IUD 0 0 0 0 0 0 0000 HU LLL L1 E 3 P N lt 17x 100mm 10000000000 10000 00000 1p gr E 7D D U D 00000 Gene Pulser Xcell Home S pombe Protocol Detail 10000 Pre set Protocols Fungal Protocols 18 3400 00000000 GP Xcell Instruction Manual 54
124. n for protocol 1 User1 2 Labguy 5 Malcom 4 Smith 5 Wesson 6 03 9 User Director 00000 User Directoryp 000000000000000000000000000000000 000000000000000000000000000000000 User 00000000000000 User Protocols USER PROTOCOLS Useri Choose location for protocol 1 CHO 2 Monkey 5 Human 4 5 6 03 10 User Protocolsq 0 0 0 00 User Protocols 0 0000000000000000000000000000000 000000000000000000000000000000000000 User 50 0000000000 00000 GP Xcell Instruction Manual 16
125. ngal 5 Fungal 6 13 4 Gene Pulser Xcell 100000 0000000000 1000000000000 00000 0000 00000 000000 msec HF Q cm 1 CHO Square wave 15 160 02 100 COS7 Square wave 20 110 02 100 3T3 Exponential decay 500 160 0 2 100 293 Square 25 110 0 2 100 HeLa Exponential decay 500 00 160 0 2 100 BHK21 Square wave 25 140 02 100 A549 Square wave 10 150 02 100 CV1 Square wave 25 100 02 100 K562 Exponential decay 1000 155 0 2 100 HL60 Square wave 25 140 02 100 J urkat Exponential decay 1000 140 0 2 100 HuT78 Square wave 25 130 02 100 E coli Exponential decay 25 200 1800 041 20 E coli Exponential decay 25 200 2500 02 20 40 E coli Exponential decay 25 200 3000 02 20 40 A tumefaciens Exponential decay 25 200 2400 01 20 P aeruginosa Exponential decay 25 200 2500 0 100 S aureus Exponential decay 25 100 2900 02 50 B cereus Exponential decay 50 200 1000 02 100 S pyogenes Exponential decay 25 200 2100 02 200 L plantarum Exponential decay 25 400 1500 02 40 S cerevisiae Exponential decay 25 200 1500 02 40 S cerevisiae Exponential decay 25 200 2500 04 80 S pombe Exponential decay 25 200 2300 02 40 C albicans Exponential decay 25 200 1500 02 40 P pastoris Exponential decay 25 200 2000 02 40 D discoideum Square wave 1 0 1000 0 4 800 2 pulses 5 sec int GP Xcell Instruction Manual 3 4 3 0000000000000
126. put 1230 1230 1000 111 1 Data output Data output Manual exponential Settings V 1230 TC mseo 1854 5 Cuvette mm 1 Data output Manual Mode Time 12 12 01 20 22 Output 1230 1854 5 Standard Protocol S pombe setting Data output 12 12 01 80 85 Sed Protocol S pombe Settings Output V 2000 2047 25 25 R Cohm 200 200 mseo 5 1 Cuvette mm 2 2 03 30 Data output O00 GP Xcell Instruction Manual 39 0000 100000000000000000000000000000000000000000000 module 100000000000000000000 Home 0 0 3200000 00000000000 Measurements 7000000 ENTERO 00 0 0 00 Measurements 3
127. r X cell 000000000000 0 C 25u 2000 V 18kV 0000000000 0 0 0 254 2000 2 5 1000 C 25u 2000 V 3 0kV 100 Gene Pulser X cell main 0 PC module D E coli 5ml 2820 000 500m L Broth 10 37 C 300rpm 100000000 10000 100000 1000000 100000 1000000 5x 10 5 MID 00000000000000 10 20 00000000000000 10000000 50070 00 0 0 0 O 4 4 4000x 1500 0 0 100 1000000000000000000000000000000 100 100000000 4 00 4000x 150 000 100000000 250m0 0000000 201 4000x gf 1500 D D U 00000 2m 0001000000000000 00 0 4 4000x gr 1500 D 0 000 0 100 11 2 1000 10 3 10 cells 6 612 00000000000 100000000 1000000000 10000000000000 0 2 00000000 1ml 5001 0 0 17 100 100000000 1000000 1 5 0 0000000 100000000 0000 204 I 02cm 00000 201 40mID 00000000000 29 0 0 0 0 0 0 DNA D W D I 0 100000000000000000100000000000000000 100000 0 0000000000000000000000000000000000000
128. references 00000 00 0000 GP Xcell Instruction Manual 34 3 10 3 0000000 100 100000 100000 390000000 100 100000000 0 100000000000000000 I User Preferences 0 00000000 0000000000 Sleep function 000000 Sleep Function 330p 00000000 Sleep Function SLEEP FUNCTION Time before entering sleep mode 5 10 20 50 60 OFF 03 37 Sleep Function 10000000000000000000 000 000000 ENTER OOO User Preferences 60 0000000000 3 11 Pulse Trach
129. rpm 130 5x 10 cells ml 081 0 85 10000 Saureus 37 00000 30000000000 10040 100000 150 00000000000000 500mg 000000000 12 000 gr 15000 LU D D UO OE BD UB DE WE E D DE C 0000000000000 500m0 0000000 1 12 000 91 IQ 0 0 0 0000 00000 0 0 000 10000000000 50m 20000000 45 GP Xcell Instruction Manual 1000000000000000 00 000000 25 0 0 30ml 401 12000x g 15 0000000000000 10000 1070000000 200 000 11500000000 4 4 12 000 g 1510 00 0000 Oakridge N 10000 11000000 2770 00000000000 1x 10 101 1570 0000 2501 00000000000000000000000000000 70 DNA 5 9 2u 90 000000 100000000 5041
130. ruginosa by electroporation Nuc Acids Res 17 10509 1989 Suzuki T Shin B C Fujikura K Matsuzaki T and Takata K Direct gene transfer into rat liver cells by in vivo electroporation FEBS Lett 425 436 1998 Thompson J R Register E Curotto J Kurtz M and Kelly R An improved protocol for the preparation of yeast cells for transformation by electroporation Yeast 14 565 1998 Trevors J T Chassy B M Dower W J and Blaschek H P Electrotransformation of bacteria by plasmid DNA in Guide to Electroporation and Electrofusion Chang D C Chassy B M Saunders J A and Sowers A E eds Academic Press Inc p 265 San Diego 1992 Willson T A and Gough N M High voltage E coli electro transformation with DNA following ligation Nuc Acids Res 16 11820 1988 Zabarovsky E R and Winberg G High efficiency electroporation of ligated DNA into bacteria Nuc Acids Res 18 5912 1990 63 GP Xcell Instruction Manual 0190 0000000 101 Gene Pulser Xcell 000000000 0000 00 0000 WxDxH 100 120 220 240VAC 50 60Hz 240W 000 0 35 0 95 61010 61326 Class main unit 3 1 30x14cm module 31x 28x 9cm PC module 31x 28x 5cm main unit 6 62kg CE module 3 08kg PC module 1 90kg 00000000000000 CE moduler
131. sa apu NOR te cer 58 RIEL tiia 59 59 811 59 82 27 RN a 59 59 ERE 60 saa k ay 61 o E ah 64 eet aan be On a una A LO E 64 Gene Pulser 0 000000000000009000800
132. tL LU DU Capacitor Measurement Capacitor measurement Capacitance XXXX Measured capacitance uF XXXX 3 33 Capacitor Measurement 3 10 00000000000 Home 000 3 20 0 0 19 10000 1000000000 User Preferences 1000 User preferences 1000 User Preferences User Preferences 1 Clock intensity Sleep function 3 34 User Preferences 33 GP Xcell Instruction Manual
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