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Team:KULeuven/11 August 2008
From 2008.igem.org
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Lab Work
Wet Lab
- We did PCR on the following ligations: J23109 + J23032, I712074 + J23032, GFP-LVA + B0015, R0062 + B0032, R0084 + J23022 and C0012 + B0015. R0084+B0032 did not gave colonies. This is the second time. Tomorrow we are going to ligate this parts again.
- We continued to construct the T7 polymerase with UmuD tag using PCR.
- We electroporated the following parts: C0060 + B0015, C0040 + B0015, J23100 + B0032.
- We MiniPrepped the liquid cultures we made last week. After that, we cut C0060, C0062, R0040 with EcoRI and SpeI. F1610 was cut with EcoRI and B0032 was cut with XbaI.
Dry Lab
Modeling
DAY OF THE MODEL REVIEW
We found a small (but important) mistake in our model: the efficiency of the rbs for the translation of LacI was set to ony 0.05 while all the other efficienties of the rbs that are incorporated in the lock3d were put on 0.167. This makes a serious difference in the final result and not in a good way: the amount of produced LuxI (and following HSL/HSL_LuxR-complex) is reduced a lot! We we're able to slightly repair this new fact by increasing the efficiency of the rbs of translation of LuxI to maximal, but this doesn't completely fix the problem. It only results in the formation of 2 to 3 molecules of ccdB in the cell which might not be lethal enough...
It seems there's been quite a big misunderstanding between mathematical modellers and biochemical modellers which has resulted in yet another mistake in the model. This time it light be more severe, because after a fast repair, the preliminary results indicate that the timer-capabilities have completely vanished...
Let's hope that the night will bring some good idea's!
Wiki
Quote of the day
“It is better to be roughly right than precisely wrong” - [J.M. Keynes]
Remarks
Multicroc is gesloten tot 27 aug!
