Hi everybody, I am trying to determine the phenotype of my transformation of pichia, I have the strains, X33 and SMD1168H, I did changes in the recommendations of the protocol of easy select pichia of thermo. The transformants were replicated in liquid YPD medium and when I get OD in log phase I adjusted to the same concentration every colony and I inoculated in MD Y MM mediums in plates. I don't have control currently and I got this plaques for example:
The criteria for select is: Mut+ strains will grow normally on both plates, while Muts strains will grow normally on the MD plate but show little or no growth on the MM plate.
I think that all my transformants cells are Muts strains they grew normally on the MD plate but show minor growth on the MM plate.
What do you think?
I also thought repeat the assay but decrease the concentration when I inoculated, in this assay I Inoculated to OD=2
In blue X33 in black SMD1168H
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