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SEPTEMBER: WEEK 3
September, 12th
E31-1, E41N-1, E42-1, E43-3 and ENTERO4C5 were diluted 1:100 in 13 falcon tubes (4ml M9 + Cm12.5 pH = 6.0). After two hours they were induced with aTc (6 ng/ml, 8 ng/ml and 100 ng/ml). Supernatants were taken at the induction time, after 1 hour, 2 hours and 4 hours.
September, 13th
Supernatants collected on September, 12th were tested, measuring 3OC6-HSL concentration with T9002-ENTERO in two differents tests.
September, 14th
T9002-ENTERO, ENTERO-RBS were diluted 1:100 in M9 + Amp + Cm12.5 respectively in 18 ml and 1 ml. After two hours a test was performed on supernatants collected on September, 13th.
As we realized that T9002-ENTERO did not produce the desired output (saturation of known induction curve occured at 5 nM, even if we expected that it was in the linear working zone of the biosensor) we decided to use BBa_T9002 for our measurements, so we inoculated it together with RBS in M9 + Amp, in order to perform tests on supernatants from cultures expressing AiiA. September, 15th
BBa_T9002 and RBS cultures were diluted 1:100 in M9 + Amp; after two hours the test on supernatants of cultures expressing AiiA enzyme at various aTc inductions started. In the afternoon the over night liquid cultures were diluted 1:100 again in order to test the remaining supernatants collected.
September, 16th
BBa_T9002 and RBS cultures were diluted 1:100 in M9 + Amp; after two hours the test on supernatants of cultures not expressing AiiA enzyme and M9 without cells at pH = 6.0 and pH = 7.0 started (SN collected on September, 14th).
September, 17thWiki update and definition of results. September, 18thWiki update and definition of results.
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Team:UNIPV-Pavia/Calendar/September/week3
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