Chemical transformation in 96 well MTP - How to make competent cells in MTPs (Dec/03/2008 )

Hi,

We would like to make transformation experiments in 96 well MTP format in E. coli. This sounds easy in that case wen you need only one strain of competent cells, but I have to transform hundred different mutant with the same expression vector. My question is that are there any good protocol for making competent cells (whether chemical or electrocompetent is irrelevant) in 96 well microtiter plate format?

A rapid, and easy method for making competent cells would be also good for this purpose. If there is I would optimize it for MTPs. I know only one rapid method which utilizes 50mM of CaCl2 for making chemical competent cells.


Using Sacharomyces there are existing methods using MTPs for chemical transfomation.

Any Idea?

It's probably pretty easy to make electrocompetent cells, which just essentially involves washing with water multiple times. I would think you could do that in deep well 96 well plates, then electroporate with a 96 well electroporator.

Since you are transforming with plasmid, you probably don't need high efficiency, so the CaCl2 preps at 10^6 to 10^7 cfu/ug probably would work just fine. They would be easy to do in deep well 96 well plates.

Thanks for answering! My ideas were almost similar to that you wrote. Probably it is not difficult to handle cells in deep well MTPs in order to make some competent cells from them.

Thank you again!