Dear people,

I have a plasmid (pCMV-miR) that contains, f1 ori, ColE1 ori as well as SV40 ori. As I thought it, my plasmid is a cosmid (Phagemid) with very low copy number.

When I transform my bacteria wit wheither empty vector or insert containing vector I always get colonies after 24 h. The more, when I pick a colony from kan+ agar and inoculate it to the kan+ LB or kan+ tryptic soy broth I got OD nearly 36h later.

I am using 50mg/l kanamycin. Do you suggest me to reduce antibody to 30mg/l ? Or is the problem caused by some problem else?

I use dh5a as carrier.

Thanks for answers