Growing of TOP10 chemically competent cells - (Jun/11/2009 )

Is there anybody who has grown chemically competent TOP10 cells from invitrogen stock successfullyat lab.

If anybody did it, would you please share with us?
I found the protocol at Wiki, but want advice from experienced persons who are doing this regularly.

Thanks,

-epigenetics-

Here is one protocol:

http://openwetware.org/wiki/TOP10_chemically_competent_cells

But I always make competent E. coli cells with RbCl-method, that has always worked for me (DH5a, S17-1, BL21).
And I think that a colleague of mine has used that protocol for a TOP10 derivative strain too...

-mastermi-

mastermi on Jun 11 2009, 12:52 PM said:

Thanks,

I have this one, Did anybody follow this (in the link) for TOP10 cells?

-epigenetics-

They're just E. coli, and will grow like any other E. coli strain. The resulting growth will not, of course, be chemically competent any more, and so you must make them so. I use a RbCl2-based method as well -- the protocol I use is available here -- and I have used it successfully on TOP10 cells, as well as on many other E. coli (and Pseudomonas) strains...

-HomeBrew-

HomeBrew on Jun 11 2009, 01:13 PM said:

thanks a lot

-epigenetics-