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Star Republic: Guide for Biologists

Transformation --- electroporation

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.