ELECTROPORATION OF HEAVILY ENCAPSULATED BACTERIA - (May/21/2008 )

HI there I have been trying unsucessfully to electroporate a heavily encapsulated bacteria. Does anyone know about any electroporation conditions thta woudl improve the efficinecy of electroporation. Thanks

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Are you getting a lot of arcing? Or is that you don't see any transformants after incubation? Also, check your settings:

I electroporate at 1.8 kV, 25 uFD, and 200 ohms. I also use 0.1cm cuvets.

When you make electrocompetent stocks of your host bacteria, make sure you wash it GOOD! Try washing with lots and lots of sterile ddH20 (not just distilled, but deionized as well) and then suspending in a solution of 10% glycerol. Try using only 20uL of bacteria in your next electroporation, and only 1uL of DNA that is in water or TE buffer. (Preferably water)

If that doesn't work, you may have to try chemical transformation.

I hope this helps!

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Just wondering if you know the capsule composition, you can try to (partially) digest it first. If it works, write a biotechniques paper

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