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Team:UNIPV-Pavia/Notebook/Week4Jun
From 2009.igem.org
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Week from June 22nd, to June 28th, 2009
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June, 22nd
- This week we planned to ligate the GFP protein generator under the control of Ptet in A4 and A6 constructs, in order to have:
- an aTc->GFP measurement system (A6-E0240 = J23100-RBS-tetR-TT-Ptet-RBS-GFP-TT) and
- a control construct that should always express GFP(A4-E0240 = RBS-tetR-TT-Ptet-RBS-GFP-TT)
- We infected 5 ml of LB + Amp with 10 ul of these glycerol stocks (to prepare samples for this week's ligations):
| K117000 | E0240 (X5) | J23118 |
| A4 | A6 |
- We incubated the 5 ml cultures overnight at 37°C, 220 rpm.
- Wiki updating.
June, 23rd
- Miniprep for: E0240 (5 samples), A4, A6, TT, J23118 and K117000
- Digestions:
| E0240(X-P) | J23101(E-S) | K117000(E-S) |
| A4(S-P) | A6(S-P) | TT(E-X) |
- The bands were cut from the gel and the following ligations were performed:
- A7 = J23118(S-P) + E0240(X-P)
- A8 = A4(S-P) + E0240(X-P)
- A9 = A6(S-P) + E0240(X-P)
- A10 = K117000(E-S) + TT(E-X)
- The ligation reactions were incubated overnight at 16°C.
June, 24th
June, 25th
June, 26th
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