Team:Brasil-SP/Notebook
From 2014.igem.org
WELCOME TO iGEM 2014!Your team has been approved and you are ready to start the iGEM season!
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Notebook | ||||||||||||
You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well. |
Experimental Methodology
For the assemblies constructuion we addopted the following method:
![](https://static.igem.org/mediawiki/2014/f/fc/Metodologia_Brasil-SP.png)
Main Assembly Map
![](https://static.igem.org/mediawiki/2014/8/87/Mapa.png)
Assemblies forms
Assembly form template
This is the template designed to help with the laboratory organization during the assemble of biological parts. If you want to use the assembly method we used you can print the form and complete it for each construction. Hope it helps :)
![](https://static.igem.org/mediawiki/2014/3/32/Assembly_form_pg3.png)
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Characterization Assemblies
Apart from the main genetic circuit we also assembled others for characterization purposes, such as the validation of the promoters and tunning of our threshold setter concentration, the QteE.
Promoter BBa_K823003
Question: Does the constitutive promoter BBa_K823003 work properly?
![](https://static.igem.org/mediawiki/2014/d/d5/KI_w.png)
Results: After the incubation period of the transformed E. coli a large portion of the colonies were glowing green. So the promoter does work. Moreover, this biobrick works on E. coli despite the fact it was designed for B. subtilis.
Promoter BBa_K143015
Question: How does the transcription caused by this promoter varies with the IPTG induction?
![](https://static.igem.org/mediawiki/2014/7/7a/KXVI_w.png)
Results:
Tunning of the QteE Threshold
Question:What are the concentration of QteE needed to hamper the LasR induction of the promoter PlasR?
This is the most difficult task of our project. Tunning the production of QteE so that we establish the correct threshold for the discretization of the Cystatin C level in serum. To attack this challenge we designed 3 circuits so that we could plot a calibration curve. In this circuits we put the transcription of the LasR and QteE under two differnt promoters, the Pveg (BBa_K823003) and PlasR (BBa_K143015).
![](https://static.igem.org/mediawiki/2014/8/86/KXV_w.png)
![](https://static.igem.org/mediawiki/2014/c/c4/KXIII_w.png)
![](https://static.igem.org/mediawiki/2014/4/43/KXIV_w.png)
Assembly Forms
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Life Inside the LAB
July
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October
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