Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.502 12/2001 Microorganism Agrobacterium tumefaciens Cell type Bacteria, gram negative Molecules injected Plasmid DNA (in water) Growth medium TB medium Washing solution Sterile, cold water Electroporation solution 10% glycerol Outgrowth medium LB medium Cuvette 1 mm gap width Reference Mersereau M., et al 1990 Gene 90 149-151

Making electrocompetent cells:

1. Grow cells at 30 C to a density of O.D.₆₀₀: 1.5-2.0.


2. Harvest by centrifugation and wash five to seven times in sterile, cold water.


3. Resuspend the cells in 10% glycerol to a final concentration of 4-6 x 10¹¹ cell/ml and store frozen at 70 C.


4. For electroporation frozen cells were thawed in ice and diluted with sterile water to a density of 1 x 10¹⁰ cells/ml prior to use.

Electroporation of cells:

1. Add 2 ng plasmid DNA to 40 l of electrocompetent cells. Homogenize by gently mixing with pipette several times.

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