I am planning to do maize protoplast assay (dual-LUC assay) with an effector construct under the estrogen inducible promoter and my question is if anyone knows when should the induction be done i.e. at what point is it best to add the estradiol, after electroporation or after the O/N incubation?
Hi Tina, I suggest consult the Sheen Protocol´s for protoplasts obtention
But you will required that you are registred with your e-mail....
http://molbio.mgh.harvard.edu/sheenweb/protocols_reg.html
When I used EtOH to induce expression I added it after electroporation when the samples are in their indicidual reaction vessels.
Hi, Tina, you should add induction agent directly to the plates with protoplasts after transfection. Do not change the medium, just add inducer to the plates with cells! Good luck!
Thank you Miguel, Yuming, Falco, and Taras, these suggestions were helpful, so just wanted to say thanks... I am not very active on RG so sorry for not replying back sooner.
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