Day 1
- pick a colony (e.g. E. coli BL21(DE3) or similar) from a recently plated transformation into ~3 mL LB with appropriate antibiotic
- incubate with shaking at 37 C overnight
Day 2
- Add 400 µL of overnight culture to a new eppendorf tube; microcentrifuge max speed for ~1 min
- Discard supernatant, but leave a drop with the cell pellet
- Add a drop of loading buffer to the inside of the eppendorf lid; add 20 µL of phenol-chloroform to the pellet and vortex
- Invert to mix in the loading buffer and microcentrifuge at max speed for 5 min
- Load 15-20 µL of the upper phase on agarose gel