So, initially I have transformed a plasmid into the clinical E.coli isolate. Now, i need to co-tranform these cells with another plasmid of choice. When i tried making the cells with plasmid 1 competent, they arent showing the desired resukts, i.e, the cells arent being competent. What may be the reason for this. Both plasmids require same antibiotic so that is not the problem here i guess.
Why dont you perform co-transformation of both plasmids together? And re-clone one of the genes in a diff plasmids so that 2 antibiotics can be used rather than 1.
Thank you so much Megha Abbey for your quick response, I will try the same.
It is going to be very hard to identify a co-transformant if you have the same antibiotic on both plasmids since you don't have a simple way to identify whether a cell has plasmid A, plasmid B or both. As Megha Abbey mentioned, the best solution is to make one of the plasmid carry a different antibiotic resistant gene. Or alternatively put both genes on the same plasmid.
While co-transormation does work well, you are again going to run into the problem of how to identify the co-transformants rather than the single transformants.
Thank you Michael J. Benedik I will work on this.
Thank you for your response.
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