pBAD/Myc - Gene Synthesis

PurificationScale-up ofExpression forPurificationUse the conditions determined in the previous section to grow and induce 50 ml of cells.This is the largest culture volume to use with the 2 ml prepacked columns included in theProBond Purification System. If you need to purify larger amounts of recombinantprotein, you may need more ProBond resin. See page 2 for ordering information. Note:Remember to use RM medium (page 26) with LMG194.1. Inoculate 2 ml of SOB or LB containing 50 µg/ml ampicillin with a singlerecombinant E. coli colony.2. Grow overnight at 37°C with shaking (225-250 rpm) to OD 600 = 1-2.3. The next day, inoculate 50 ml of SOB or LB containing 50 µg/ml ampicillin with1 ml of the overnight culture.4. Grow the culture at 37°C with vigorous shaking to an OD 600 = ~0.5 (the cells shouldbe in mid-log phase).5. Add the optimal amount of L-arabinose to induce expression.6. Grow at 37°C with shaking until the optimal time point is reached. Harvest the cellsby centrifugation (3000 x g for 10 minutes at +4°C).7. At this point, you may proceed directly to purification (see ProBond PurificationSystem manual) or store at -80°C for future use.PurificationFor help with purification of your recombinant protein, please refer to the ProBond Purification System manual.If you are using another type of resin, please refer to the manufacturer's recommendations.21