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Transformation problems - (Mar/17/2006 )

I have a problem in transforming DH5 alpha electrocompetent cells. I tried to transform uncut vector alone without any insert still it did not give me any colonies.

All transformations were done by electroporation at 1700 volts.

Thanks.

-SHYR-

hmmmm....I would make new comp cells. Do you have protocol that has worked well for you in the past? would you like to post your protocol so that we may be able to help you find the problem, for both making the cells and performing the transformation?

-aimikins-

i agree with aimikins, after having eliminated problems of antibiotics too concentred when preparation of plate, or mistake in antibiotic choosen.
Finally you may try with an other plasmid to eliminate possibility of pb in your own plasmid.

-fred_33-