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How to make electrocompetent cells without refrigerated centrifugue - (Sep/12/2006 )

Hi

The refrigerated centrifugue at the lab are broken down, and I really need to do E.coli HD5 alpha electrocompetent cells.

Do you known any protocol to do that without refrigerated centrifugue and without cold room.

Thanks

-djv0022-

do you have an electrical outlet in your fridge?

we have a 'deli-style' refrigerator in our lab, that comes with outlets. it's easy enough to put a microfuge in there. it might be a pain for you, but perhaps you could make your comp cells in a very small batch for now, till you get better access to a cold centrifuge

-aimikins-

If you are centrifuge at lower than 1000rpm, then you can put the eppendorf tubes in a 50ml tube filled with ice. Remeber to balance the 50ml tube.

Hope this may help.

-Minnie Mouse-

Thanks for the idea aimikins, but all the refrigerators at lab are full, no one with space for microcentrifugue.

Minnie thats a good idea, but the centrifuge boken down are the only for 50 ml falcon tubes.

My boss tell me about to do the protocol in ten eppendorf tubes on microcentrifugue without refrigeration, for CdCl competent cells I dont kwown how efficient are that protocol in that way.

But I also need electroporate Bacillus thuringiensis, so I will wait for the repair.

Thanks

-djv0022-