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DH5 keep kicking out my vector - (Dec/31/2007 )

I´m working with a lentiviral vector. I cloned several different genes into it successfully, but now when I want to make new DNA out of the glycerin stocks, I can´t isolate the DNA. I tried different Midi and Mini prep techniques, I also tried transforming the DNA back into the DH5 and starting with fresh from the plate bacteria, also tried different medium and different amp concentrations and growing temps..... nothing helps.
I don´t know what else to do. I´d be eternally grateful for any help,
thank you and Happy New Year:)



for lentivirus, we use stbl3 cells to grow them in. Also I wouldnt use the DH5-alpha, as they might not prevent recombination and this could alter virus production. You could also try SURE cells.


Thank you very much for your answer, I´m trying to transform the vector into STbl3 tonight.
Again thanks, you saved me a lot of worry smile.gif