Induction of EPI300 Bacterial Cells

Culture EPI300 cells carrying an inducible vector orvernight (in 5-10 ml selective medium).

1. Subculture 1:10 or 1:100 into 0.05-0.5 liter fresh selective medium supplemented with 0.04% Arabinose or 1X Lucigen's Induction Solution (CCIS125).

2. Grow (with rotation) at 30°C or 37°C (you'll get more DNA at 37°C) for few more hours (around 6 hours for a 1:10 dilution, more for a 1:100). Ideally, monitor OD600 for best results. You can follow the recommendations in the NucleoBond XtraBAC Kit manual, that recommends a total pellet size of 2.8 g, achievable, for example, by harvesting 250 ml bacteria at OD600=6 or 500 ml bacteria at OD600=3. Importantly, when measuring OD600 on the nanodrop:

   1. Blank with LB

   2. Dilute your bacterial cultures 1:5 or 1:10 as the nanodrop measurements saturates at OD600>1.

   3. Back-calculate your undiluted OD600 by multiplying by the dilution factor.

3. Prep DNA using your preferred method.

NOTE: if your DNA is transformed into an E. coli strain that is not EPI300, copy number amplification will not happen!!