Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.510 12/2001 Microorganism Corynebacterium glutamicum Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium LBG (LB with 0.5% glucose) supplemented with 2.5% glycine and isonicotinic acid hydrazid (4 mg/ml) Washing solution 15% glycerol (v/v) Electroporation solution 15% glycerol (v/v) Outgrowth medium LBG medium (LB with 0.5% glucose) Cuvette 2 mm gap width Reference Haynes J. and Britz M. 1990 Journal of General Microbiology 136 255-263 Making electrocompetent cells:

1. Grow cells in LBG medium at 30 C and shaking at 200 rpm to an O.D.₆₀₀ of 0.15 to 0.25. 2. Harvest by centrifugation. 3. Wash in one culture volume of 15% glycerol. 4. Resuspend cells in 0.002 culture volumes of 15% glycerol (number of cells: approx. 2.5 x 10¹⁰ cells/ml).
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1. Corynebacterium glutamicum