Team:Paris Bettencourt/Notebook/TB-ception/Wednesday 16th October.html

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TB-ception

Tuesday 16th October


We want to investigate the stability of the protein TDMH at different pH.
To do this, I prepared buffers at different pH using monosodium phosphate and disodium phosphate.
We used a calculator (http://home.fuse.net/clymer/buffers/phos2.html) to have an idea of how much miligrams should we add of each to get the right pH. Buffer strenght was 10mM.

We prepared a range of pH and checked them using a pH meter.

The pH were: 4,54 ; 5,15; 6,43; 7,36; 8,25 and 8,84

pH lower than 4,5 and higher than 8,8 couldn't be reached (because of the pKa as it is a weak acid).

We wanted to test another pH around 3, so we add HCl to reach pH 3.

The buffer were made with osmosed water and have been autoclaved after the pH measurement.


We also made glycerol stock of E.coli (the one with 3 plasmids carrying TDMH,LLO and RFP genes)from the overnight as described in the protocol.