Edited by Chang Zhu
Cells growing in liquid medium
1. Grow cells to late log phase.
2. Pellet cells and wash with sterile dH2O.
3. Wash with 1 M sorbitol.
4. Resuspend cells in ice-cold 1 M sorbitol (1-5x 109 cells/ml).
Cells on plates
1a. Scape about 20 ml cells from
a plate.
2a. wash with sterile dH2O.
3a. Wash with 1 M sorbitol.
4a. Resuspend cells in 40 ml ice-cold 1 M sorbitol.
Electroporation
5. Add 0.1-1 mg DNA and incubate on ice for 5 minutes.
6. Transfer to a 2 mm electroporation cuvette.
7. Electroporate with a field strength of 7.5-9 kV/cm (1500-1800 V).
8. Add 0.25 ml 1 M sorbitol.
9. Plate on selective medium.
10. Transformants appear in 4-6 days at 30 °C.
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