Team:Paris Bettencourt/Notebook/TB-ception/Thursday 1st August.html



Thursday 1st August

We want to see if 1 mM solution of IPTG induces the over expression in our strain. We prepared 4 liquid cultures (no antibiotics added):

• bl21 (DE3) + pET21b strain induced with 1 mM IPTG
• uninduced bl21 (DE3) + pET21b strain
• untransformed bl21 (DE3) + 1 mM IPTG
• untransformed bl21 (DE3)

10 mL of culture had 500 µL of the overnight culture (O.D.600 was set to 0.2) and 100 µL of 100 mM IPTG. We measured the O.D. of the cultures (600 nm) for time 0 and after 6 h. We also diluted our cultures to calculate the CFU/mL (for time 0 10-1 and 10-2 dilutions were prepared, and 100 µL of each dilution was plated on LBA plates; for 6 h 10-3, 10-4 and 10-5 for the induced culture and 10-7, 10-8 and 10-9 for all the others, 10 µL of each were plated). We also saved 2 mL for each group in the fridge.