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One Shot® ccdB Survival™-T1R Chemically Competent Cells: Gateway cloning????


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#1 Rits

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Posted 30 September 2009 - 03:52 AM

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

#2 phage434

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Posted 30 September 2009 - 04:04 AM

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

#3 Rits

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Posted 30 September 2009 - 04:41 AM

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



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

#4 phage434

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Posted 30 September 2009 - 12:36 PM

Here's my favorite protocol:
http://openwetware.o...competent_cells

#5 maoniuniu

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Posted 04 September 2010 - 07:57 PM

Here's my favorite protocol:
http://openwetware.o...competent_cells

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.

#6 phage434

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Posted 06 September 2010 - 08:14 AM

We have used that protocol with DB3.1 successfully.




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