Is it possible that A. tumefaciens A136 encounter some type of Mutation after serial culturing? if not then why it's not giving positive result even for positive control in Quorum sensing investigation?
Hi,
Yes. it is possible that A136 mutated and lost its insert. I suggest try to obtain a new strain. Additionally, check your protocol? Which media do you use to culture your strain?
Sergey
@Christina. I am trying to get quantitative analysis at first, just to know either it have AHL activity or not? can you please asset me about the short protocol? I'll be really thankful.
regards
@Sergey Dobretsov, Sir I am using Marin Agar on which both A136 and my test strains grows well but not responding to X-Gal even to positive control. sir my protocols are as follow
1. Culturing of Reporter strains using Marine Agar.
2. Culturing of Positive and test strains > incubating them for 24/48 hours > culturing them with repoter strain A136 in T manner> incubatin them for 24/48 Hours at 28 °C > adding X-Gal > Cheking for QS activity after 12 hours then after 24 hours then after 36 hours then after 48 hours.
Sir is there any thing wrong in this protocol? if yes kindly guied me.
Respectful Regards,
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