ALPHA SYNUCLEIN WT EXPRESSION

Materials:

Transformed BL21 cells containing Alpha Synuclein pet23a Vector

Kanamycin Stock 10mg/ml (10ug/mL working concentration)

250mL LB KAN; 500mL LB KAN

1M IPTG Stock

Protocol:

1) Made 250mL O/N culture Inoculated with a scratch from a glycerol stock. Or
picked colony from kanamycin plate. Incubate @37C with shaking overnight.
2) Inoculated 500L LB KAN with 50ml of O/N culture. Incubate with shaking
@37C.
3) Add 1mM IPTG when OD600 of the culture is between 0.6 and 0.8 (takes
approximately 1.5 2 hrs).
4) Induce for 2.5 hrs @37C with shaking.
5) Harvested induced cells. 2 X 250ml pellets in 50mL conical tubes. Store at
minus 70C.