“I have two questions , thw first one is how can I increase growth of competent cells DB3.1 in LB btoth media containing amp / kanamycin The second , I use vector BACMAM pCMV dest which contains lethal gene CCdb and needs LR mix reaction , in the protocol it's mentioned that LR mix reaction transformed in DH10BAc , so can I use DH5alpha or DH10 B instead of DH10Bac .. ? ”
Hi Dina,
for your first question you could try using TB media instead of LB to increase the growth. This media has more nutrients than standard LB.
Your second really needs more information. If you are putting your gene into the pCMV-dest vector then you can use DH5alpha to propagate and purify the plasmid. If you are going to make virus with it then you have to use the DH10Bac cells. If you are trying to propagate just the empty pCMV-Dest vector then you have to use Survivor cells from ThermoFisher as the vector has the CCDB gene that will kill other strains of E.coli.
I hope that helps.
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