Hi. I am trying to transfer RK2 plasmid from donor to recipient only. Does anybody give me an idea how I can distinguish transconjugants and recipients in the medium, they will have same genotype.Thanks
If the RK2 plasmid carried certain antibiotic resistance gene or other selection marker, you could add certain antibiotic in the selection medium. The recipient always has some antibiotic resistance genotype, such as the E. coli J53, which was a rifampicin resistant strain, so the transconjugants should be selected in medium with both rifampicin (depends on the recipient you use ) and the RK2 plasmid resistance antibiotic.
The problem is counterselection against the donor. The recipient has an antibiotic resistence which is lacking in the donor. You must look up which strains you used and select for the plasmid and against the donor.
Thanks for the answers. My problem is that transconjugant cells can grow in recipient cell medium. I think there should be something else in recipient cells or transconjugant cells to select them only.
If u dont have double antibiotic selection as suggested above than other option is to have a differential media with one antibiotic which your recipient strain is senitive and select based on the donor and transconjugant differentiation on the differential media and once you get the colonies....do multiple time subculture to get the pure colony of transconjugant. I did with E.coli and rhizobium on congo red agar. It works but require lots of effort and time. I hope it will work for you
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