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Team:KULeuven/NotebookDaily
From 2011.igem.org
tuesday 5th of July
Cell culture medium and agar plates containing antibiotics (ampicillin, kanamycin, tetracyclin and chloramphenicol) were prepared. A precul- ture of competent cells was made (protocol competent cells).
wednesday 6th of July
Competent cells were prepared according to the CaCl2 method.
thursday 7th of July
Biobricks were transformed in competent cells. List of biobricks:
1. pLux-CI promotor BBa K091107
2. pLac/Mnt Hybrid Promoter BBa K091104
3. cspA Promoter BBa K328001
4. luxI Bba C0061
5. luxR Bba C0062
6. pconst-RBS-MelA Bb K193602
7. CrtEBI Bba K274100
8. CI repressor BBa R0051
9. Mnt repressor BBa C0072
10. Colicin E2 operon without SOS promotor BBa K131009
11. ribolock3c BBa J23031
12. ribokey3c Bba J23008
13. ribosome binding site BBa B0034
14. terminator BBa B0015
15. OmpA Bba K103006
16. TEV cleavage site Bba K128002
17. GFP Generator BBa E0240
These transformed cells were grown on agar plates containing the cor- rect antibiotic for selection.
friday 8th of July
We observed some colonies on the plates, so the transformation of bio- bricks worked.
tuesday 12th of July
Colonies were picked and grown in liquid medium overnight at 37°C. Another transformation of competent cells with new biobricks was done:
1. BAD promotor BBa I13453 2. ribokey3d BBa J23066
The Cu promotor was replaced by the BAD promotor.
wednesday 13th of July
Minipreparations of the 17 biobricks were performed according to the protocol of the kit (promega miniprep kit)(Fig.1).
thursday 14th of July
New cultures for minipreparations were prepared since the DNA con- centrations of the previous miniprep were too low to go on with restric- tion. A new transformation of the competent cells was performed: luxI BBa C0261 (this will be our new luxI instead of Bba C0061 which we transformed more early).
friday 15th of July
6ml cultures were miniprepped and after DNA gel electrophoresis the concentration of the di�erent biobricks was estimated. The DNA con- centrations were higher than the �rst miniprep and su�cient to proceed with restriction. (Fig.2)
IMAGE
Fig.1: Miniprep 13/07: L, A= pLux-CI promotor, B= pLac/Mnt Hybrid Promoter, D= Ribolock-Coilicin E2 Dnase, E= ribolock3c, F= ribokey3d, G= GFP Generator, H= CrtEBI, I= CI repressor, J= ribosome binding site, K1= terminator, K2= pconst-RBS-MelA, Z= CspA Promoter, 1= luxI, 2= luxR , 3= Mnt repressor, 4= OmpA , 5= TEV cleavage site, L
IMAGE
Fig.2: Miniprep 15/07: labelling is the same as in Fig.1 except for K= pconst-RBS-MelA and Y= terminator.
wednesday 13th of July
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wednesday 13th of July
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wednesday 13th of July
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wednesday 13th of July
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friday 22th of July
After reformatting the laptop, it took windows 2 hours to update! Installation of maya, an amazing 3D-visualising program, provided by iGEM sponsor the mathworks. We checked the price and normally it costs $3,251! Time to watch tutorials... But first an evening dinner with our team.
monday 25th of July
Importation of 3D- structures of icenucleating protein and anti-freezeprotein in maya: check! Found some cool video made in maya. So, it should be possible to make one, but how long will it takes us?
tuesday 26th of July
Working in simbiology.
wednesday 27th of July
We made a simulation for a hybrid promotor lac_lux (BBa_R091100) which is almost synthesized in the wet lab coupled to GFP. Values of parameters are estimated, a recurring theme in kinetic modelling?
thursday 28th of July
Today we brainstormed on how we could make a molecular movie about are project.
