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Bifidobacterium animalis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.538 04/2002 Microorganism Bifidobacterium animalis Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium MRS broth with 0.05% cysteine.HCl and 0.5 M sucrose (final concentrations) Washing solution 0.5 M sucrose Electroporation solution 0.5 M sucrose, 1 mM ammonium citrate, pH 6 Outgrowth medium MRS broth with 0.05% cysteine.HCl and 0.5 M sucrose (final concentrations) Cuvette 1 mm gap width Reference Argnani, A. et al 1996 Microbiology 142 109-114 Making electrocompetent cells:

1. Cultivate cells by using an overnight culture to inoculate fresh medium. Grow cells overnight at 37 C. Dilute this culture 1:25 in fresh medium and cultivate at 37 C until an O.D.695 of 0.2. Chill bacteria on ice. 2. Harvest by centrifugation. 3. Wash twice with 0.5 M sucrose 4. Resuspend in about 1/250 of the original culture volume
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