Team:Paris Bettencourt/Notebook/TB-ception/Saturday 26th October.html
From 2013.igem.org
TB-ception
ASDFTuesday 16th October
We repeated the pH study using the plate reader and the OD this time.
Same controls and conditions, the only difference is the protein extraction.
We decided that we will just overexpressed E.coli (10mM IPTG as previously) and collect the supernatant.
We centrifuged 5minutes at 4000 rpm and took the supernatant. Then, the supernatant was filter sterilized
We add 200 microliters of M.smegmatis (OD 0,05) resuspended in the appropriate buffer and 20 microliters of E.coli (OD 0,5).
To know precisely the initial number of bacteria, we made CFU for each pH (resuspended in a different buffer).
We launched the plate reader for 24 hours.