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Team:Paris Bettencourt/Notebook/TB-ception/Sunday 15th September.html
From 2013.igem.org
TB-ception
ASDFMonday 15th September
• Liquid culture for TDMH+pSB1C3.
• Colony PCR for confirming TDMH+pSB1C3 with biobrick verification primers.
• A new setup for testing if E. coli kills M. smegmatis in the macrophages: 2 wells with M. smegmatis, 1 well with M. smeg + E. coli uninduced, 1 well with only E. coli, 1 well with M. smeg + E. coli induced. First we infect the macropheges with the liquid culture of M. smeg GFP - we set the final O.D. of the culture in the well to 0.03 (100 µL of the liquid culture with O.D. 0.3 to 900 µL of RPMI with attached macrophages. After 2 h of incubation in the CO2 incubator we wash the cells with PBS 10 times and we add E. coli induced or uninduced (induced overnight with 10 µL IPTG).
