My miniprep gives high yield but maxiprep gives no plasmid? Am I losing my plasmid because it is too large or unstable? How can the bacteria grow without the plasmid?
I am trying to produce stock of an 11kb lentivector in stellar cells (i.e. suitable cells for lenti vector). My miniprep had a pretty good yield - 15ug from just 1.5ml culture. The 1.5ml for the mini came from a 5ml O/N (14hrs) culture started from a well isolated single colony on a plate that was less than a day old. I setup two 100ml maxi cultures - seeded one with 100ul and another with 1ml from the 5ml miniculture. The amplicillin was 100ug/ml in both mini and maxi. Maxi was grown O/N for 14 hrs. Both maxi cultures had good sized pellets but no plasmid yield?