One Shot® ccdB Survival™-T1R Chemically Competent Cells: Gateway cloning???? - problems using competent cells for destination vectors in gateway clon (Sep/30/2009 )

Hi all....

Has/is anyone using these new ccDB survival competent cells from Invitrogen ; which have replaced the preexisiting DB3.1 cells for cloning of vectors with ccDB cassette???

I have tried these cells for the transformations with my destination vectors; and have tried a couple of different batches sent by the company but with no success or only a few colonies per plate as compared to DB3.1 cells where transformations worked beautifully over the plates.

I am wondering if anyone has experience of using these cells and if these cells are working fine for them.

Unfortunately Invitrogen do not supply the DB3.1 cells anymore.


Please help, any information or suggestion in this regard will be highly appreciated


Rits

You could simply make some of your existing DB3.1 cells competent and use them. (Cheaper, also).

phage434 on Sep 30 2009, 01:04 PM said:

Unfortunately I don't have any DB3.1 cells myself. But possibly could ask around if someone could let me have some.

If so, Can you please guiide me on how to make own competent cells (I heard there are tricks in these commercially available cells though!!???!!)

Rits

Here's my favorite protocol:
http://openwetware.org/wiki/TOP10_chemically_competent_cells

phage434 on Wed Sep 30 20:36:58 2009 said:

Hi,recently I want to make some competent cells using DB3.1 strain,so whether your protocal are entirely appropriate for DB3.1? or need some changes?
Thanks.

We have used that protocol with DB3.1 successfully.