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Nocardia corallina

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.526 01/2002 Microorganism Nocardia corallina Cell type Bacteria, gram positive Molecules injected Plasmid DNA (in TE buffer) Growth medium N2 medium (20 g/l glucose, 2 g/l yeast extract, 4 g/l beef extract, 6 g/l tryptone, 2 g/l NaCl, 10 g/l glycine) Washing solution Distilled water Electroporation solution 0.3 M sucrose, 15% glycerol Outgrowth medium N2 medium (without antibiotics) Cuvette 2 mm gap width Reference Yao W., et al 1994 Current Microbiology 29 223-227 Making electrocompetent cells:

1. Inoculate a fresh overnight culture of bacteria into N2 medium. Grow cells at 28 C to an O.D.600 of 0.7. Harvest by centrifugation.. 2. Wash twice with _ volume of distilled water. 3. Resuspend in a final volume of 1/20 of the original volume of electroporation solution.

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