Hi Everyone,
I am currently trying to standardize a protocol for Agrobacterium mediated gene transfer in plants. As a crucial step, induction of vir genes in Agro is mandatory and the composition specifies MES buffer pH5.6. As of now, we have only Tris- Cl buffer in our lab and MES is falling expensive. Can I use Tris- Cl pH 5.6 instead? Does it make a difference while using it to induce vir genes in Agro?
Thanks!
MES and acetosyringone are commonly used to induce vir gene. You can use acetosyringone if MES is not available. I would not recommend Tris-Cl as a substitute for MES. Agro strain/plant being used are also important which optimizing protocol.
Good luck
1. MES is a buffer.
2. MES buffer's useful pH range is from pH 5.5-6.7 (see attached chart). Tris-HCl's useful buffer range is from pH 7.0-9.0 (see Sigma website: type in 'Trisma-HCL', Trisma is Sigma's Trademark for Tris). So, if you want to prepare a buffer with pH 5.6, MES should be the choice.
You can use Phosphate buffer instead of MES. But keep buffer pH in range of 5.6-5.8. Vir gene induction by Acetosyringone takes place at acidic pH of 5.6. A good protocol in this regard is provided by SB Gelvin and you can avoid MES in second buffer but maintain pH.
https://www.bio.purdue.edu/people/faculty/gelvin/gelvinweb/AgroInductionProtocol.html
Thank you everyone for your valuable answers.. Kamal Kumar, thank you very much for the idea as I can atleast try using phosphate buffer till I purchase MES..
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