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Kluyveromyces lactis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.516 12/2001 Microorganism Kluyveromyces lactis AWJ137 Cell type Yeast Molecules injected DNA Growth medium Not given Washing solution YPD, 10 mM DTT, 40 mM HEPES (pH 8.0) Electroporation solution 2.5 mM Tris/HCl, 1 mM MgCl2, (pH 8.0) Outgrowth medium YPD with 1 M sorbitol Cuvette 4 mm gap width Reference Prof. Friedhelm Meinhardt Universitt Mnster Corrensstr. 3 D-48149 Mnster
e-mail: meinhar@uni-muenster.de Making electrocompetent cells:

1. Grow a 50 ml cell culture of Kluyveromyces lactis to a density of 5 x 107 cells/ml. 2. Harvest by centrifugation (5 min., at 4,000 x g) 3. Resuspend cells in 5 ml washing solution and incubate for 5 min. 4. Wash twice with electroporation solution and resuspend in 0.25 ml of the same solution. Incubate for 1 hour on ice.
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