Team:UNIPV-Pavia/Calendar/August/week4

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UNIPV TEAM 2011

March
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April
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May
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30 31

June
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July
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August
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22 23 24 25 26 27 28
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September
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October
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31

AUGUST: WEEK 4

August, 22nd

Streak of E17-2, E18-2, J101-31, J101-E5 and ENTERO-4C5 on LB agar + Cm12.5 plate to test Plux promoter with different RBSs.
E24-1, E25-2, E26-1, E27-2 and E41N-1 purified DNA samples were sent to BMR Genomics for sequencing.
250 ml of M9 glycerol supplemented minimal medium were prepared.
Inoculum of T9002-ENTERO and ENTERO-RBS to test supernatants collected on August, 20th to test AiiA enzyme efficiency.

August, 23rd

T9002-ENTERO and ENTERO-RBS were diluted 1:500 in a final volume of 1 ml of M9 + Cm12.5.
Three colonies of E17-2, E18-2, J101-31, J101-E5 and ENTERO-4C5 were picked and inoculated in 1 ml of M9 + Cm12.5.
E19-2, E 20-2, J101-E7, J101-4C5 and ENTEOR-4C5 were streaked on a LB agar + Cm12.5 plate.
Unfortunately TECAN test could not be performed as the instrument did not work!
31 LB agar + Cm34 plates were prepared.
Last week sequencing were ready: E28-1 has not the desired sequence, while E37-2, E38-1, E39-1, E40-2 and E42-1 were correct.
Ptet-RBS30-luxI ligation was done again from E36 and E2:

Ligation Name Vector Vector volume (μl) Insert Insert volume (μl) Buffer (μl) T4 Ligase (μl)
E43 E36 (S-P) 3.5 E2 (X-P) 4.5 1 1