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):

• 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:

• 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

• We resuspended R0011 (=pLac) and K112808 (=lysis actuator from T4 phage) 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.
• Team meeting
• We received sequencing results for:
  • K131009 - BioBrick was correct, but prefix/suffix sequences had a missing nucleotide...We noticed this problem even in the sequencing performed by MIT. We will document it on K131009 page. Anyway, the restriction sites were not corrupted.
  • A3 - correct! (long part, but lacZ had already been successfully tested)
  • A4 - correct!
  • A5 - correct! (long part, but lacZ had already been successfully tested)
  • A6 - correct!

June, 25th

June, 26th

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