I am transforming a recombinant plasmid into top10 e.coli strain, to replicate more plasmid so I can use it in LAB bacteria.
And several observations makes me believe that there could be some leaky expression of the protein even in top 10 cell that causing some toxicity.
When I change the incubation temperature to 30 instead of 37, I manage to get positive colonies.
I am trying to transform another recombinant plasmid that express bigger protein, the temperature strategy alone doesn't solve the leaky expression problem anymore. I am wondering if I should grow it at even lower temperature. Or is there other tricks I may try to solve the toxicity issue?