Team:UNIPV-Pavia/Notebook/Week4Jun

From 2009.igem.org

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*Miniprep for: E0240 (5 samples), A4, A6, TT, J23118 and K117000
*Miniprep for: E0240 (5 samples), A4, A6, TT, J23118 and K117000
*Digestions:
*Digestions:

Revision as of 22:05, 24 June 2009

EthanolPVanimation.gif

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Week from June 22nd, to June 28th, 2009

Previous Week Next Week

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.

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June, 23rd

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  • 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

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  • We resuspended R0011 (=pLac) and K112808 from the iGEM plates with 15 ul of RNAse free water.
  • Trasformations in Top10 bacteria were done for: A7, A8, A9, A10, K112808 and R0011.

June, 25th

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June, 26th

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