I did streak plate from glycerol stock. Cultures were grown on plate and in LB, they grew well initially. But when I reinoculated glycerol stock after 3 months on LB and LA media their is no sign of growth.
I prepared fresh LB and LA with appropriate antibiotics (Kan + rif).
How can I solve this problem?
Hi. I agree with Thakku. YEB medium is a very good alternative. I also recommend this publication by Wise et al. It deals generally with the best media for managing Agrobacterium in the lab.
Wise A.A., Liu Z., Binns A.N. (2006) Culture and Maintenance of Agrobacterium Strains. In: Wang K. (eds) Agrobacterium Protocols. Methods in Molecular Biology, vol 343. Humana Press. https://doi.org/10.1385/1-59745-130-4:3
https://link.springer.com/protocol/10.1385/1-59745-130-4:3#aboutcontent
YEB is the best medium to grow Agrobacterium LBA4404 strain and tries to keep the temperature around 26 to 28 degrees. Use Rif concentration 10 mg/L and Kan 50 mg/L.
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