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Differentiating Ecoli from Agrobacteria quickly.


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#1 nimrod337

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Posted 30 September 2010 - 09:10 AM

Ok, In my infinite wisdom as a young scientist, I took very poor notes. (since been altered :) ) And I had cloned my gene of interest into T-easy, then to get it to a binary vector for transforming plants (pCambia) I had to cut it out of T-easy, put it in pbluscript, then cut it from that and insert it into Pcambia. Then transform Agrobacteria to use in my plant transforming. So I ended up making glycerol stocks of the Pcambia transformed Agro/ecoli... and I made a nice note of the insert, where it was from, what resistance it had, what vector I used and all that. However, I forgot to write down if the stocks were made from the transformed Ecoli, or the transformed agrobacteria. (whoops.) So now, much later, I need to use the agrobacteria to transform some new plants, and I need to make sure I am going to transform them, not just inject them with ecoli.
So, I'm wondering if there is a quick way to screen colonies or cultures of bacteria to tell if it is Agro or Ecoli? They grow readily from glycerol stocks into overnight cultures of LB broth, which is a little disconcerning. (I've heard you need to plate up Agrobacteria before overnighting it or it won't readily multiply in liquid broth. Any suggestions? I really would like to avoid ordering some specific primers to check this, but if anyone knows of any, I'd rather be safe than sorry. Thanks,

-A

#2 phage434

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Posted 30 September 2010 - 02:25 PM

Grow a culture, miniprep, and then transform into agrobacterium. You won't have to worry.

#3 nimrod337

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Posted 01 October 2010 - 08:52 AM

Hmmm... Nice idea. I can't believe I hadn't thought of that. Thanks.

-A




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