I want to confirm is there any antibiotic resistant Lb agar plate is requires for streaking of rosetta cells for rosetta comp. cell preparation OR I can use only Lb agar plate for this purpose.
I will use this streaked plate for rosetta competent cell preparation.
I also want to know is there any difference in BL21 and rosetta competent cells preparation?
Hi there,
Rosetta as a resident plamid encoding rare tRNAs. To ensure the presence of the plasmid, chloramphenicol has to be added to media. In absence of this antibiotic the plasmid would be lost during culture. So the only difference in competent cell preparation is the addition of chlorampenicol during the culture. As a consequence you can't use CamR plasmid for expression in Rosetta cells.
Thank you linger, I want to know which antibiotic resistant plate should I use for streaking of rosetta for preparation of rosetta competent cells
You should use chloramphenicol in your LB agar. Recommended concentration is 34 μg/ml.
Actually as Dominique said, Rosetta can express codons rarely used in E. coli. These rare codons are encoded by a compatible chloramphenicol-resistant plasmid. If you don't use chloramphenicol, Rosetta will lose this plasmid and you will lose the rare-codon advantage.
I hope it's clear now!
Good luck!
Rosetta have many strains with different antibiotic resistant
see which is your strain and prepare plate accordingly
ex,
Rosetta gami need kan+cam+tet
Rosetta origami need kan+tet+cam
Rosetta (E3) plysS need only cam
good luck
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