Team:UNIPV-Pavia/Calendar/September/week3

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UNIPV TEAM 2011

March
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September
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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.
E37-2, E38-1, E39-1, E40-2 and ENTERO4C5 were inoculated in 1 ml M9 + Cm12.5 and grown ON.

September, 13th

Supernatants collected on September, 12th were tested, measuring 3OC6-HSL concentration with T9002-ENTERO in two differents tests.
1:100 dilution of E37-2, E38-1, E39-1, E40-2 and ENTERO4C5 in 4ml M9 + Cm12.5 and two hours growth, after which cultures were induced with aTc; addition of 3OC6-HSL 100 nM after one hour and collection of supernatants at t = 0h, t = 1 h, t = 2 h and t = 4 h (time refers to 3OC6-HSL addition).
Inoculum of E44 for plasmid purification in 5 ml LB + Cm34; inoculum of T9002-ENTERO and ENTERO-RBS for 3OC6-HSL concentration measurement; ENTERO4C5 was inoculated in M9 + Cm12.5 at pH = 6.0 and pH = 7.0 in order to detect 3OC6-HSL degradation in culutres not expressing AiiA.

September, 14th

September, 15th

September, 16th

September, 17th

September, 18th