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Team:UNIPV-Pavia/Notebook/Week4Jun
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== <html><font class="dayw_style">June, 24th</font></html> == | == <html><font class="dayw_style">June, 24th</font></html> == | ||
Revision as of 23:45, 23 June 2009
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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.
- We transformed in Top10 bacteria with the plasmid with J23118-B0034(RBS)-C0012(LacI)-B0015(TT) (gift from Bologna iGEM team) and plated in LB+amp plates.
- Wiki updating.
June, 23rd
- Miniprep for: E0240 (5 samples), A4, A6, TT, J23118 and K117000
- Digestions and ligations:
- A7=J23118(S-P) + E0240(X-P)
- A8=A4(S-P) + E0240(X-P)
- A9=A6(S-P) + E0240(X-P)
- A10=TT(E-X) + K117000(E-S)
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- Colonies grew in the LB+amp plates with the J23118-RBS-lacI-TT sample, we inoculated 5 ml in LB+amp o/n at 37°C.
June, 24th
June, 25th
June, 26th
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