Hi all
I have been doing this tobacco transformation using leaf disc as explant for quite some time, but i am facing so many problems. Please tissue culture experts help me out.
I have always marked that after washing the leaves cocultivated with agrobacterium, I grow them in selection medium in MS+ BAP+NAA medium with kanamycin (100 mg/l)and cefatoxime (500 mg/l) as antibiotics but I find constant agrobacterium overgrowth for weeks together even if i transfer to fresh selection media.
Please help me as i am not able to understand the situation.
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tamarine
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Posted 17 May 2012 - 04:59 PM
Hi there,
Do you measure the OD of Agrobacterium?
Some points that you have to concerned with are: Adjust the OD of Agrobacterium just on exactly written in the protocol, place the cocultivation dishes in the temperature not over than 25-26 degree Celcius, increase the antibiotic concentration if it still didnt work (but, it could poise to the explant)
Good luck
Do you measure the OD of Agrobacterium?
Some points that you have to concerned with are: Adjust the OD of Agrobacterium just on exactly written in the protocol, place the cocultivation dishes in the temperature not over than 25-26 degree Celcius, increase the antibiotic concentration if it still didnt work (but, it could poise to the explant)
Good luck
