August, 29th
T9002-ENTERo and ENTERO-RBS were diluted 1:500 in M9 + Amp + Cm12.5 13 ml and 1 ml volume respectively. After six hour growth they were aliquoted in 96-well microplate (190 μl cultures) with 10 μl of supernatants collected on August, 27th and 10 μl of 3OC6-HSL.
5 flasks were prepared with 29.6 ml H2O, 200 μl glycerol 80 %, 8 ml M9 salts 5 x and their pH was adjusted to 5.9, 6.3, 6.7, 6.9 and 7.2; they were autoclaved in order to prepare M9 glycerol supplemented minimal medium.
E43-3 DNA was digested with Ecori and PstI restriction nucleases in order to screen the part length by gel-electrophoresis; it showed the correct band.
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