Team:Paris Bettencourt/Notebook/TB-ception/Monday 12th August.html



Monday 12th August

In order to grow M. smegmatis we should add Tween 80 (0.2 % / v.v) into the LB media (it stops the cells from forming clumps). In some papers we found that Tween 80 could inhibit the growth of E. coli, so we did a growth curve to check that. 500 μL of BL21 + pET21b culture (O.D. 600 nm = 1.00) was added to 10 mL of LB supplemented with different concentrations of Tween 80 – 0.2, 0.1 and 0.05 %. There were two sets of these cultures – one was induced with IPTG and the other one was not. O.D. was measured after 3 and 6 h. At these time periods cultures were also diluted to calculate the CFU. No growth inhibition was observed, so we can use Tween 80 in our E.coli – M. smegmatis co-cultures. We have also made some plates with malachite green (40, 4 and 0.4 μg/mL) and we have plated E. coli on them.