Editing Protein Purification using Magnetic Beads (section)

=== 1. Grow Bacteria Overnight in 2 ml of Autoinduction Medium Using 24-well Plates ===

Cells: lac-inducible BL21 (DE3) Rosetta 2 cells<br>

Sterilise the 24-well plates by microwaving.<br>

<br> Inoculate 1.5 ml of autoinduction medium with freshly grown bacterial colonies or frozen glycerol stocks. <br>

Usually inoculate 2 wells per clone with 2 individual cloned colonies.<br>

Reserve 4 wells for positive (His-MBP) and 1 negative control (transformed with non-expressing plasmid)<br>

Check for sufficient sterilization of the plate by leaving one well blank for the medium-only control.<br>

Grow the cells for 18 hr at 37 °C and shaking at 250 rpm. <br>

Autoinduction medium contains a mixture of glucose and lactose. The bacteria initially feed on glucose and then begin to use lactose, which also induces the expression of the recombinant proteins.<br>