BioRad Electroporator
Gene Pulser Xcell
Contact: Debby Walthall
Rm: NSC 338
Tel: (404) 413-5363

General Warnings:
Every time you use the electroporator, you MUST sign the log book.
Balance all tubes very carefully.
Make sure everything (cuvettes, tubes, cells) is VERY COLD.
No salt in DNA.

1. Keep cells cold on ice. You can either buy or make your own competent cells. Aliquot cells and store at –70oC. Good for ~6 months. Thaw only 1 time. Thaw on ice. Cuvettes and DNA on ice.
2. No salt in DNA mix.
3. The amount of DNA and competent cells depends on the cell type. See manual for details.
4. Put DNA and competent cells in the cuvette. Make sure there aren’t any bubbles.
5. Turn Electroporator on (right side of Main unit).
6. Open pod with tab. Put in cuvette. The cuvette has metal on 2 sides – line up metal sides with metal in pod. Close pod.
7. Scroll through choices with arrows. When you get to the choice you want, select Enter.
8. Preset Protocols. There are 9 pre-set bacterial protocols, 6 pre-set Fungal protocols and 12 pre-set mammalian protocols.
a. From the home screen, Select 4 or use the arrows to scroll to Preset Protocols. Press Enter to go to the Preset protocols screen.
b. Select 1-3 or use the arrow keys to highlight either Bacterial, Fungal or Mammalian Preset Protocols. Press Enter.
c. Using the keypad or the arrow keys, scroll through the list of names. For Bacterial and Mammalian protocols, use the right and left arrow keys to toggle between the 2 columns. When the protocol you want is highlighted, press Enter. Check out the Protocol Details to make sure the electroporation parameters are what you want. You can change the parameters for pulse and you don’t have to save the change (for a one time pulse).
d. Enter
9. “P” flashing in lower right hand corner – ready to Pulse.
10. Punch red button to pulse. The LCD display will blank and then say “Pulsing”. When the pulse is done, a tone will sound and the pulse measurements will be displayed on the Protocol Results screen
11. Check time constant. It should be ~5 msec. If too high – problem.
12. Use the Left and Right arrow keys to toggle between the Protocol Results screen and the last Protocol Detail screen.
13. With the Protocol Detail screen on display, another pulse can be delivered using the same pulse parameters. To change the pulse conditions, press Enter. The cursor appears in the voltage parameter value. Make any changes that you need. Use the up and down arrows to move cursor.
14. New program. Go to # 5 from the home screen or move cursor to User Protocols. You can also modify a pre-set protocol and save it under User protocols. For instructions for doing these 2 - please use manual.

This information is given as a guide to the facilities and instrumentation available in the DNA/Protein Core facility at Georgia State University. If you have any concerns or thoughts about the content of this website please contact: John Houghton (404) 413-5390

 

 


 

 

2d analysis facscanto image-quant