pET21 contains ampicillin gene as the selectable marker and Rosetta cells require Cloramphenicol because of the pRARE plasmid? What antibiotic should I use then?
Dear Amruta
do your mean rosetta (DE3) strain?
of course you can use pet21 with this strain.
you have to use both antibiotics ( ampicillin 100mg/l and chloroanfenicol 34mg/l).
just take in consideration that Rosetta growth much lower than standard bl21(de3)
good luck
Manuele
Thanks Manuele Martinelli . Thank you so much for your reply. Does that mean after the transformation of the pET vector in the host, I should plate the cells on LB media containing both Ampicillin and Chloramphenicol?
Dear Amruta
yes, to be sure that both plasmid are retained into the cells you have to maintain both antibiotic in all the steps, plating, pre-culture and culture.
good luck
Manuele
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