( ...)Azotobacter vinelandii

Multiporator / Electroporator 2510 Transformation Protocol

Protocol No. 4308 915.503 04/2002

Microorganism Azotobacter vinelandii Cell type Bacteria, gram negative Molecules injected Plasmid DNA Growth medium Nitrogen-free medium Washing solution Ice-cold 10% sterile glycerol Electroporation solution Ice-cold 10% sterile glycerol Outgrowth medium Azotobacter growth (AG) medium Cuvette 1 mm gap width Reference Kornyi, P. et al 1998 Research in Microbiology 149 361-372

Making electrocompetent cells:

1. Cultivate cells with vigorous shaking at 30 C to an O.D.₆₂₀ of 0.4-0.5.


2. Harvest by centrifugation at 5,000 rpm for 10 min at 0 C.


3. Wash with the original culture volume of ice-cold 10% glycerol.


4. Repeat this step three times using a half and a quarter of the original volume, and finally 2-4 ml of the glycerol solution (7.0 x 10⁷ cells/ml final cell concentration).


5. Freeze 40 l aliquots in liquid nitrogen and store at 70
   '"/>

Source:


Page: All 1 2
Related biology technology :

1. Azotobacter vinelandii