I have AGL1 agrobacterium, it's not competent. i need to do it competent. any recommended protocols. i tried the TSS we use for E.coli but not working. also i tried the cacl2 protocol and don't get the +ve colonies in transfromation.
thanks
Here is my Agro competent cell protocol, I use it for AGL1 and GV3101. Usually, I make them about once a year or so, they store quite well at -80. Just be patient with the outgrowing/shaking step and watch the incubator temperature so it doesn't get too hot. Good luck!
Hi Mohamed,
we use a simple protocol similar to Amy's. It works for most agro strains we use, except for LBA4404. There, we use another one, which I can also share if you want.
With all agro strains we use LB medium supplemented with 0.2g/L MgSO4 (=LB+ in the protocol), which reduces the clumping of cells (somewhat).
Good luck,
Klaus
For Agrobacterium I usually use electro competent cells. The protocol is easier and transformation of large plant binary vector is easy with electroporation. I am sure you may find some electro cuvette and instruments at your or nearby department. All best
yes, I did electro-transformation for agrobacterium many times before. I agree with you that electrotransformation is most suited for Agrobacterium. the point is we don't have the machine and not available around. thanks
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